vero e6 cells  (ATCC)


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  • 99
    Name:
    VERO C1008 Vero 76 clone E6 Vero E6
    Description:

    Catalog Number:
    crl-1586
    Price:
    None
    Applications:
    VERO C1008 exhibits some degree of contact inhibition after forming a monolayer and is therefore useful in growing slow replicating viruses.
    Host:
    Cercopithecus aethiops
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    Structured Review

    ATCC vero e6 cells
    Stability of the GFP reporter gene expressed by rLASV-GFP. Cells were exposed to rLASV-GFP (MOI = 0.01). At 72 h PI, TCS were collected (passage 1, P1), and virus titers were determined by plaque assay. Fresh <t>Vero</t> ( a ) or Vero-E6 cells ( b ) were exposed to with TCS from P1 (MOI = 0.01). This process was serially repeated throughout P10. GFP expression (green) was measured by high-content imaging. Nuclei were stained with DAPI (blue). Viral RNA was extracted from TCS of P1 to P10 to amplify DNA fragments containing an GFP-P2A and a part of the NP ORF using RT-PCR. Images are representative field images of individual wells. Bar, 200 µm.

    https://www.bioz.com/result/vero e6 cells/product/ATCC
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    vero e6 cells - by Bioz Stars, 2020-11
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    Images

    1) Product Images from "Recombinant Lassa Virus Expressing Green Fluorescent Protein as a Tool for High-Throughput Drug Screens and Neutralizing Antibody Assays"

    Article Title: Recombinant Lassa Virus Expressing Green Fluorescent Protein as a Tool for High-Throughput Drug Screens and Neutralizing Antibody Assays

    Journal: Viruses

    doi: 10.3390/v10110655

    Stability of the GFP reporter gene expressed by rLASV-GFP. Cells were exposed to rLASV-GFP (MOI = 0.01). At 72 h PI, TCS were collected (passage 1, P1), and virus titers were determined by plaque assay. Fresh Vero ( a ) or Vero-E6 cells ( b ) were exposed to with TCS from P1 (MOI = 0.01). This process was serially repeated throughout P10. GFP expression (green) was measured by high-content imaging. Nuclei were stained with DAPI (blue). Viral RNA was extracted from TCS of P1 to P10 to amplify DNA fragments containing an GFP-P2A and a part of the NP ORF using RT-PCR. Images are representative field images of individual wells. Bar, 200 µm.
    Figure Legend Snippet: Stability of the GFP reporter gene expressed by rLASV-GFP. Cells were exposed to rLASV-GFP (MOI = 0.01). At 72 h PI, TCS were collected (passage 1, P1), and virus titers were determined by plaque assay. Fresh Vero ( a ) or Vero-E6 cells ( b ) were exposed to with TCS from P1 (MOI = 0.01). This process was serially repeated throughout P10. GFP expression (green) was measured by high-content imaging. Nuclei were stained with DAPI (blue). Viral RNA was extracted from TCS of P1 to P10 to amplify DNA fragments containing an GFP-P2A and a part of the NP ORF using RT-PCR. Images are representative field images of individual wells. Bar, 200 µm.

    Techniques Used: Plaque Assay, Expressing, Imaging, Staining, Reverse Transcription Polymerase Chain Reaction

    Antiviral drug evaluation based on recombinant LASV expressing GFP (rLASV-GFP). ( a ) Vero E6 cells were pretreated with drugs at the indicated concentrations and then exposed to rLASV-WT or rLASV-GFP (MOI = 0.1) in the presence of the drugs. Viral titers in TCS at 48 h PI were determined by plaque assay. Values represent the means ± standard deviations of triplicate samples. ( b ) Infectivity of rLASV-GFP in A549, HeLa, Huh7, and Vero E6 cells at the indicated MOIs at 24, 48, and 72h PI as determined by GFP-expression using high-content imaging. ( c ) Effect of favipiravir and ribavirin on rLASV-GFP multiplication at 48 (orange filled circles) and 72 h PI (green filled squares). Cells were exposed to rLASV-GFP (MOI = 0.1) and treated with various concentrations of favipiravir or ribavirin. The percentage of GFP-positive cells was determined at 48 h or 72 h PI. ( d ) Half-maximal effective concentrations (EC 50 ) of favipiravir and ribavirin to inhibit rLASV-GFP infection in four cell types at 48 and 72 h PI.
    Figure Legend Snippet: Antiviral drug evaluation based on recombinant LASV expressing GFP (rLASV-GFP). ( a ) Vero E6 cells were pretreated with drugs at the indicated concentrations and then exposed to rLASV-WT or rLASV-GFP (MOI = 0.1) in the presence of the drugs. Viral titers in TCS at 48 h PI were determined by plaque assay. Values represent the means ± standard deviations of triplicate samples. ( b ) Infectivity of rLASV-GFP in A549, HeLa, Huh7, and Vero E6 cells at the indicated MOIs at 24, 48, and 72h PI as determined by GFP-expression using high-content imaging. ( c ) Effect of favipiravir and ribavirin on rLASV-GFP multiplication at 48 (orange filled circles) and 72 h PI (green filled squares). Cells were exposed to rLASV-GFP (MOI = 0.1) and treated with various concentrations of favipiravir or ribavirin. The percentage of GFP-positive cells was determined at 48 h or 72 h PI. ( d ) Half-maximal effective concentrations (EC 50 ) of favipiravir and ribavirin to inhibit rLASV-GFP infection in four cell types at 48 and 72 h PI.

    Techniques Used: Recombinant, Expressing, Plaque Assay, Infection, Imaging

    2) Product Images from "Characterization and pathogenicity of a novel mammalian orthoreovirus from wild short-nosed fruit bats"

    Article Title: Characterization and pathogenicity of a novel mammalian orthoreovirus from wild short-nosed fruit bats

    Journal: Infection, Genetics and Evolution

    doi: 10.1016/j.meegid.2016.05.039

    (A) Indirect immunofluorescence detection of MRV in Vero E6 cells infected with strain B/03. The cells were fixed with 4% paraformaldehyde, blocked with 1% BSA, washed, and incubated with mouse anti-MRV (T3D) antibody. The cells were then incubated with FITC-labeled goat anti-mouse IgG secondary antibody. (B) Electron micrographs. a: Negative staining of cell culture supernatant. Non-enveloped reoviral-like particles with double-layered capsid structure were observed (diameter = approximately 70 nm). b: Ultra-thin sections of infected Vero E6 cells displayed typical contrast-rich virus particles, organized as paracrystalline structures within the cytosol. (C) SDS-PAGE demonstrating electrophoresis pattern. L, M and S represent large, medium and small segments, respectively. dsRNA segments were separated by electrophoresis in 8% ( w / v ) polyacrylamide slab gels.
    Figure Legend Snippet: (A) Indirect immunofluorescence detection of MRV in Vero E6 cells infected with strain B/03. The cells were fixed with 4% paraformaldehyde, blocked with 1% BSA, washed, and incubated with mouse anti-MRV (T3D) antibody. The cells were then incubated with FITC-labeled goat anti-mouse IgG secondary antibody. (B) Electron micrographs. a: Negative staining of cell culture supernatant. Non-enveloped reoviral-like particles with double-layered capsid structure were observed (diameter = approximately 70 nm). b: Ultra-thin sections of infected Vero E6 cells displayed typical contrast-rich virus particles, organized as paracrystalline structures within the cytosol. (C) SDS-PAGE demonstrating electrophoresis pattern. L, M and S represent large, medium and small segments, respectively. dsRNA segments were separated by electrophoresis in 8% ( w / v ) polyacrylamide slab gels.

    Techniques Used: Immunofluorescence, Infection, Incubation, Labeling, Negative Staining, Cell Culture, SDS Page, Electrophoresis

    3) Product Images from "A Small interfering RNA lead targeting RNA-dependent RNA-polymerase effectively inhibit the SARS-CoV-2 infection in Golden Syrian hamster and Rhesus macaque"

    Article Title: A Small interfering RNA lead targeting RNA-dependent RNA-polymerase effectively inhibit the SARS-CoV-2 infection in Golden Syrian hamster and Rhesus macaque

    Journal: bioRxiv

    doi: 10.1101/2020.07.07.190967

    siRNA No. 14 inhibition of SARS-CoV-2 cytopathicity in Vero E6 cells (CPE assay and EC 50 ). Vero E6 cells were infected with SARS-CoV-2 and incubated for 2 days. (a) Mock-siRNA (100 nM). (b) 5 nM. (c) 10 nM. (d) 20 nM. (e) 30 nM. (f) 40 nM. (g) 50 nM. (h) 60 nM. (i) 70 nM. (j) 80 nM. (k) 90 nM. (l) 100 nM. (m) EC 50 of siRNA No. 14 using qRT-PCR.
    Figure Legend Snippet: siRNA No. 14 inhibition of SARS-CoV-2 cytopathicity in Vero E6 cells (CPE assay and EC 50 ). Vero E6 cells were infected with SARS-CoV-2 and incubated for 2 days. (a) Mock-siRNA (100 nM). (b) 5 nM. (c) 10 nM. (d) 20 nM. (e) 30 nM. (f) 40 nM. (g) 50 nM. (h) 60 nM. (i) 70 nM. (j) 80 nM. (k) 90 nM. (l) 100 nM. (m) EC 50 of siRNA No. 14 using qRT-PCR.

    Techniques Used: Inhibition, Infection, Incubation, Quantitative RT-PCR

    4) Product Images from "Multivalent Presentation of Antihantavirus Peptides on Nanoparticles Enhances Infection Blockade "

    Article Title: Multivalent Presentation of Antihantavirus Peptides on Nanoparticles Enhances Infection Blockade

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.01415-07

    Focus reduction assay comparing the abilities of monovalent peptides versus those of multivalent peptide-coated nanoparticles to block SNV infection of Vero E6 cells. Prior to being added to confluent monolayers of Vero E6 cells, SNV was treated with
    Figure Legend Snippet: Focus reduction assay comparing the abilities of monovalent peptides versus those of multivalent peptide-coated nanoparticles to block SNV infection of Vero E6 cells. Prior to being added to confluent monolayers of Vero E6 cells, SNV was treated with

    Techniques Used: Blocking Assay, Infection

    5) Product Images from "Analysis of N-Linked Glycosylation of Hantaan Virus Glycoproteins and the Role of Oligosaccharide Side Chains in Protein Folding and Intracellular Trafficking"

    Article Title: Analysis of N-Linked Glycosylation of Hantaan Virus Glycoproteins and the Role of Oligosaccharide Side Chains in Protein Folding and Intracellular Trafficking

    Journal: Journal of Virology

    doi: 10.1128/JVI.78.10.5414-5422.2004

    Immunoreactivity of wild type (wt) and mutant HTNV glycoproteins with individual anti-Gn and -Gc MAbs. Vero E6 cells were infected with vTF7-3 followed by transfection with wt HTNV M cDNA (pGEM-HTNM) or glycosylation site mutant cDNA as indicated. Cells were then labeled with 50 μCi of [ 35 S]methionine for 15 h and immunoprecipitated with a pair of anti-Gn and anti-Gc MAbs. The resulting immunoprecipitates were analyzed by SDS-10% PAGE under reducing conditions.
    Figure Legend Snippet: Immunoreactivity of wild type (wt) and mutant HTNV glycoproteins with individual anti-Gn and -Gc MAbs. Vero E6 cells were infected with vTF7-3 followed by transfection with wt HTNV M cDNA (pGEM-HTNM) or glycosylation site mutant cDNA as indicated. Cells were then labeled with 50 μCi of [ 35 S]methionine for 15 h and immunoprecipitated with a pair of anti-Gn and anti-Gc MAbs. The resulting immunoprecipitates were analyzed by SDS-10% PAGE under reducing conditions.

    Techniques Used: Mutagenesis, Infection, Transfection, Labeling, Immunoprecipitation, Polyacrylamide Gel Electrophoresis

    Biochemical analyses of N-glycan chains on HTNV glycoproteins Gn and Gc. (A) DNJ blocks N-glycan trimming of Gn and Gc. Vero E6 cells were infected with vT-HTN M and radiolabeled with 50 μCi of [ 35 S]methionine for 15 h in the absence (lanes 1 to 3) or presence (lanes 4 to 6) of 2 mM DNJ, and cell lysates were immunoprecipitated with anti-Gn and -Gc MAbs. The resulting precipitates were subjected to digestion with endo H and PNGase F for 20 h as indicated and analyzed by SDS-10% PAGE under reducing conditions. The deglycosylated forms of Gn and Gc are marked dGn and dGc, respectively. (B) endo H and PNGase F digestion of HTNV glycoproteins labeled with [ 3 H]mannose. Vero E6 cells infected with vT-HTN M were labeled with 200 μCi of [ 3 H]mannose for 4 h, and cell lysates were reacted with anti-Gn and -Gc MAbs, followed by digestion with endo H and PNGase F. The gel was exposed for 60 days. (C) Effect of tunicamycin on glycosylation of HTNV glycoproteins. Vero E6 cells were infected with vTF7-3 and then mock transfected (lane 3) or transfected with HTNV M-segment cDNA tagged with FLAG. Cells were labeled with 50 μCi of [ 35 S]methionine for 15 h in the absence or presence of tunicamycin (2 μg/ml) and immunoprecipitated with anti-Gn and -Gc MAbs (lanes 1 to 3) or anti-FLAG and anti-Gc antibodies (lanes 4 to 7). The immunoprecipitates were subjected to digestion with endo H or PNGase F as indicated.
    Figure Legend Snippet: Biochemical analyses of N-glycan chains on HTNV glycoproteins Gn and Gc. (A) DNJ blocks N-glycan trimming of Gn and Gc. Vero E6 cells were infected with vT-HTN M and radiolabeled with 50 μCi of [ 35 S]methionine for 15 h in the absence (lanes 1 to 3) or presence (lanes 4 to 6) of 2 mM DNJ, and cell lysates were immunoprecipitated with anti-Gn and -Gc MAbs. The resulting precipitates were subjected to digestion with endo H and PNGase F for 20 h as indicated and analyzed by SDS-10% PAGE under reducing conditions. The deglycosylated forms of Gn and Gc are marked dGn and dGc, respectively. (B) endo H and PNGase F digestion of HTNV glycoproteins labeled with [ 3 H]mannose. Vero E6 cells infected with vT-HTN M were labeled with 200 μCi of [ 3 H]mannose for 4 h, and cell lysates were reacted with anti-Gn and -Gc MAbs, followed by digestion with endo H and PNGase F. The gel was exposed for 60 days. (C) Effect of tunicamycin on glycosylation of HTNV glycoproteins. Vero E6 cells were infected with vTF7-3 and then mock transfected (lane 3) or transfected with HTNV M-segment cDNA tagged with FLAG. Cells were labeled with 50 μCi of [ 35 S]methionine for 15 h in the absence or presence of tunicamycin (2 μg/ml) and immunoprecipitated with anti-Gn and -Gc MAbs (lanes 1 to 3) or anti-FLAG and anti-Gc antibodies (lanes 4 to 7). The immunoprecipitates were subjected to digestion with endo H or PNGase F as indicated.

    Techniques Used: Infection, Immunoprecipitation, Polyacrylamide Gel Electrophoresis, Labeling, Transfection

    Association of HTNV glycoproteins with CNX and CRT. Vero E6 cells were infected with vTF7-3 followed by transfection with wild-type or N glycosylation site mutant HTNV M cDNAs and then labeled for 30 min with 50 μCi of [ 35 S]methionine in the absence (A and B, lanes 1, 3, 5, and 7; C, lanes 1 to 4) or presence (A and B, lanes 2, 4, 6, and 8) of DNJ. Cell lysates were divided into three aliquots for immunoprecipitation with anti-Gn/Gc (A), anti-CNX (B), or anti-CRT (C) antibodies. The resulting immunoprecipitates were analyzed by SDS-10% PAGE under reducing conditions.
    Figure Legend Snippet: Association of HTNV glycoproteins with CNX and CRT. Vero E6 cells were infected with vTF7-3 followed by transfection with wild-type or N glycosylation site mutant HTNV M cDNAs and then labeled for 30 min with 50 μCi of [ 35 S]methionine in the absence (A and B, lanes 1, 3, 5, and 7; C, lanes 1 to 4) or presence (A and B, lanes 2, 4, 6, and 8) of DNJ. Cell lysates were divided into three aliquots for immunoprecipitation with anti-Gn/Gc (A), anti-CNX (B), or anti-CRT (C) antibodies. The resulting immunoprecipitates were analyzed by SDS-10% PAGE under reducing conditions.

    Techniques Used: Infection, Transfection, Mutagenesis, Labeling, Immunoprecipitation, Polyacrylamide Gel Electrophoresis

    Expression of single N glycosylation site mutants of HTNV glycoproteins. Vero E6 cells were infected with vTF7-3 followed by transfection with wild type (Wt) or mutated HTNV M cDNA. Cells were labeled with 50 μCi of [ 35 S]methionine for 15 h and then immunoprecipitated with anti-Gn and -Gc MAbs. Immunoprecipitates were analyzed by SDS-10% PAGE under reducing conditions.
    Figure Legend Snippet: Expression of single N glycosylation site mutants of HTNV glycoproteins. Vero E6 cells were infected with vTF7-3 followed by transfection with wild type (Wt) or mutated HTNV M cDNA. Cells were labeled with 50 μCi of [ 35 S]methionine for 15 h and then immunoprecipitated with anti-Gn and -Gc MAbs. Immunoprecipitates were analyzed by SDS-10% PAGE under reducing conditions.

    Techniques Used: Expressing, Infection, Transfection, Labeling, Immunoprecipitation, Polyacrylamide Gel Electrophoresis

    Pulse-chase analysis of wild type (wt) and three N glycosylation site mutant glycoproteins. Vero E6 cells were infected with vTF7-3 followed by transfection with wt or mutant cDNA, labeled for 20 min with 80 μCi of [ 35 S]methionine (200 μCi/ml), and then incubated with excess unlabeled methionine for the indicated times. Cell lysates were immunoprecipitated with anti-Gn MAb 16D2 and two anti-Gc MAbs, 11E10 and HCO2. The resulting immunoprecipitates were analyzed by SDS-10% PAGE under reducing conditions (A) or by SDS-8% PAGE under nonreducing conditions (B).
    Figure Legend Snippet: Pulse-chase analysis of wild type (wt) and three N glycosylation site mutant glycoproteins. Vero E6 cells were infected with vTF7-3 followed by transfection with wt or mutant cDNA, labeled for 20 min with 80 μCi of [ 35 S]methionine (200 μCi/ml), and then incubated with excess unlabeled methionine for the indicated times. Cell lysates were immunoprecipitated with anti-Gn MAb 16D2 and two anti-Gc MAbs, 11E10 and HCO2. The resulting immunoprecipitates were analyzed by SDS-10% PAGE under reducing conditions (A) or by SDS-8% PAGE under nonreducing conditions (B).

    Techniques Used: Pulse Chase, Mutagenesis, Infection, Transfection, Labeling, Incubation, Immunoprecipitation, Polyacrylamide Gel Electrophoresis

    6) Product Images from "Synthesis of 1-?-D-ribofuranosyl-3-ethynyl-[1,2,4]triazole and its in vitro and in vivo efficacy against Hantavirus"

    Article Title: Synthesis of 1-?-D-ribofuranosyl-3-ethynyl-[1,2,4]triazole and its in vitro and in vivo efficacy against Hantavirus

    Journal:

    doi: 10.1016/j.antiviral.2008.02.003

    Metabolism of ETAR in Vero E6 cells
    Figure Legend Snippet: Metabolism of ETAR in Vero E6 cells

    Techniques Used:

    Dose response of ETAR in Vero E6 cells infected with HTNV or ANDV
    Figure Legend Snippet: Dose response of ETAR in Vero E6 cells infected with HTNV or ANDV

    Techniques Used: Infection

    Effect of ETAR on intracellular GTP levels in Vero E6 cells
    Figure Legend Snippet: Effect of ETAR on intracellular GTP levels in Vero E6 cells

    Techniques Used:

    7) Product Images from "Methyltransferase-like 3 modulates severe acute respiratory syndrome coronavirus-2 RNA N6-methyladenosine modification and replication"

    Article Title: Methyltransferase-like 3 modulates severe acute respiratory syndrome coronavirus-2 RNA N6-methyladenosine modification and replication

    Journal: bioRxiv

    doi: 10.1101/2020.10.14.338558

    SARS-CoV-2 genomic RNA harbored m 6 A modifications. (A) MeRIP and northern blotting. RNAs from virus-infected Vero-E6 cells were incubated with IgG or anti-m 6 A-specific antibodies as indicated. Immunoprecipitated RNAs were resolved on 1% agarose gels containing 2.2 M formaldehyde and transferred to Hybond-N + membranes, followed by RNA signal detection with SARS-CoV-2-specific probes spanning from nt 28,274 to nt 29,870. (B–D) MeRIP-Seq. Fragmented total RNAs from SARS-CoV-2-infected Vero-E6 cells were subjected to IP with anti-m 6 A-specific antibodies, followed by next-generation sequencing. Methylation coverage of the full-length SARS-CoV-2 RNA is shown. Representative of n = 3 determinations. (E) Table of m 6 A peak regions. The five peak regions are shown.
    Figure Legend Snippet: SARS-CoV-2 genomic RNA harbored m 6 A modifications. (A) MeRIP and northern blotting. RNAs from virus-infected Vero-E6 cells were incubated with IgG or anti-m 6 A-specific antibodies as indicated. Immunoprecipitated RNAs were resolved on 1% agarose gels containing 2.2 M formaldehyde and transferred to Hybond-N + membranes, followed by RNA signal detection with SARS-CoV-2-specific probes spanning from nt 28,274 to nt 29,870. (B–D) MeRIP-Seq. Fragmented total RNAs from SARS-CoV-2-infected Vero-E6 cells were subjected to IP with anti-m 6 A-specific antibodies, followed by next-generation sequencing. Methylation coverage of the full-length SARS-CoV-2 RNA is shown. Representative of n = 3 determinations. (E) Table of m 6 A peak regions. The five peak regions are shown.

    Techniques Used: Northern Blot, Infection, Incubation, Immunoprecipitation, Next-Generation Sequencing, Methylation

    METTL3 catalyzed the m 6 A modification of SARS-CoV-2. (A and D) Western blotting. METTL3 was knocked down by shRNA (D) or overexpressed (A) in Vero-E6 cells. The expression of METTL3 was checked using anti-METTL3 (A and D) or anti-Flag antibodies (A), as indicated. Vector-transfected cells were used as a control. (B) Formaldehyde-RIP qRT-PCR. Cell lysates from formaldehyde-crosslinking were subjected to IP with IgG or anti-Flag antibodies. qRT-PCR was performed to quantify SARS-CoV-2 RNA. IgG was used as a negative control. Unpaired Student’s t -tests were performed, and data are presented as means ± standard errors of the means (n = 3). ** P ≤ 0.01. (C and E) MeRIP-qPCR. RNA was extracted from SARS-CoV-2-infected Vero-E6 cells in which METTL3 was overexpressed (C) or knocked down by shRNA (E). Me-RIP was performed, and SARS-CoV-2 RNA was quantified by qRT-PCR. Unpaired Student’s t -tests were performed, and data are presented as means ± standard errors of the means (n = 3). ** P ≤ 0.01. (F) MeRIP and northern blotting. RNAs were harvested from SARS-CoV-2-infected Vero-E6 cells in which METTL3 was knocked down by shRNA. MeRIP and northern blotting were performed. (G) MeRIP-Seq. Total RNA was isolated from SARS-CoV-2-infected Vero E6 cells in which METTL3 was knocked down.
    Figure Legend Snippet: METTL3 catalyzed the m 6 A modification of SARS-CoV-2. (A and D) Western blotting. METTL3 was knocked down by shRNA (D) or overexpressed (A) in Vero-E6 cells. The expression of METTL3 was checked using anti-METTL3 (A and D) or anti-Flag antibodies (A), as indicated. Vector-transfected cells were used as a control. (B) Formaldehyde-RIP qRT-PCR. Cell lysates from formaldehyde-crosslinking were subjected to IP with IgG or anti-Flag antibodies. qRT-PCR was performed to quantify SARS-CoV-2 RNA. IgG was used as a negative control. Unpaired Student’s t -tests were performed, and data are presented as means ± standard errors of the means (n = 3). ** P ≤ 0.01. (C and E) MeRIP-qPCR. RNA was extracted from SARS-CoV-2-infected Vero-E6 cells in which METTL3 was overexpressed (C) or knocked down by shRNA (E). Me-RIP was performed, and SARS-CoV-2 RNA was quantified by qRT-PCR. Unpaired Student’s t -tests were performed, and data are presented as means ± standard errors of the means (n = 3). ** P ≤ 0.01. (F) MeRIP and northern blotting. RNAs were harvested from SARS-CoV-2-infected Vero-E6 cells in which METTL3 was knocked down by shRNA. MeRIP and northern blotting were performed. (G) MeRIP-Seq. Total RNA was isolated from SARS-CoV-2-infected Vero E6 cells in which METTL3 was knocked down.

    Techniques Used: Modification, Western Blot, shRNA, Expressing, Plasmid Preparation, Transfection, Quantitative RT-PCR, Negative Control, Real-time Polymerase Chain Reaction, Infection, Northern Blot, Isolation

    METTL3 regulated SARS-CoV-2 replication. (A and B) Western blotting. METTL3 and FTO was knocked down in Vero-E6 cells by shRNA. The expression of METTL3 protein and viral NP was detected by western blotting with specific antibodies. (C–F) qRT-PCR. Total RNA was isolated from SARS-CoV-2-infected Vero-E6 cells in which METTL3 and FTO was knocked down by shRNA as indicated. SARS-CoV-2 RNA was quantified using qRT-PCR with specific primers targeting N and RdRp genes. GAPDH was used as a control. Unpaired Student’s t -tests were performed. Data are presented as means ± SEMs (n = 3). * P ≤ 0.05.
    Figure Legend Snippet: METTL3 regulated SARS-CoV-2 replication. (A and B) Western blotting. METTL3 and FTO was knocked down in Vero-E6 cells by shRNA. The expression of METTL3 protein and viral NP was detected by western blotting with specific antibodies. (C–F) qRT-PCR. Total RNA was isolated from SARS-CoV-2-infected Vero-E6 cells in which METTL3 and FTO was knocked down by shRNA as indicated. SARS-CoV-2 RNA was quantified using qRT-PCR with specific primers targeting N and RdRp genes. GAPDH was used as a control. Unpaired Student’s t -tests were performed. Data are presented as means ± SEMs (n = 3). * P ≤ 0.05.

    Techniques Used: Western Blot, shRNA, Expressing, Quantitative RT-PCR, Isolation, Infection

    SARS-CoV-2 infection influenced the expression patterns of m 6 A-related proteins. Vero-E6 cells infected with SARS-CoV-2 (MOI = 0.01) were harvested at 24 and 48 hpi. Western blotting was performed with antibodies as indicated. GAPDH was used as a loading control. (B–F) Confocal microscopy images of SARS-CoV-2-or mock-infected Vero-E6 cells. The nucleus (blue) and virus protein NP (green) were labeled with Hoechst and anti-NP1-specific antibodies, respectively. The methyltransferases and demethylases were stained with antibodies as indicated. Scale bars, 5 μm.
    Figure Legend Snippet: SARS-CoV-2 infection influenced the expression patterns of m 6 A-related proteins. Vero-E6 cells infected with SARS-CoV-2 (MOI = 0.01) were harvested at 24 and 48 hpi. Western blotting was performed with antibodies as indicated. GAPDH was used as a loading control. (B–F) Confocal microscopy images of SARS-CoV-2-or mock-infected Vero-E6 cells. The nucleus (blue) and virus protein NP (green) were labeled with Hoechst and anti-NP1-specific antibodies, respectively. The methyltransferases and demethylases were stained with antibodies as indicated. Scale bars, 5 μm.

    Techniques Used: Infection, Expressing, Western Blot, Confocal Microscopy, Labeling, Staining

    RdRp expression regulated the sumoylation and ubiquitination of METTL3. (A) RNA expression of host methyltransferases. Total RNA was harvested from SARS-CoV-2-infected Vero-E6 cells every 2 h, as indicated. The mRNAs were separated and subjected to next-generation sequencing. RNA levels of host methyltransferases were normalized according to the sequencing reads. (B) Sumoylation assay. METTL3 was overexpressed in HEK293T cells by transfection with pMETTL3, followed by transfection with pFlag-RdRp, pHA-SUMO-1, and pMyc-Ubc9. IP and immunoblot analyses were performed using the indicated antibodies for the sumoylation assay. (C–E) Ubiquitination assay. HEK293T cells were transfected with pFlag-RdRp, pHA-Ubi, pHA-K48, and pHA-K63 after METTL3 overexpression. IP and immunoblot analyses were performed using the indicated antibodies.
    Figure Legend Snippet: RdRp expression regulated the sumoylation and ubiquitination of METTL3. (A) RNA expression of host methyltransferases. Total RNA was harvested from SARS-CoV-2-infected Vero-E6 cells every 2 h, as indicated. The mRNAs were separated and subjected to next-generation sequencing. RNA levels of host methyltransferases were normalized according to the sequencing reads. (B) Sumoylation assay. METTL3 was overexpressed in HEK293T cells by transfection with pMETTL3, followed by transfection with pFlag-RdRp, pHA-SUMO-1, and pMyc-Ubc9. IP and immunoblot analyses were performed using the indicated antibodies for the sumoylation assay. (C–E) Ubiquitination assay. HEK293T cells were transfected with pFlag-RdRp, pHA-Ubi, pHA-K48, and pHA-K63 after METTL3 overexpression. IP and immunoblot analyses were performed using the indicated antibodies.

    Techniques Used: Expressing, RNA Expression, Infection, Next-Generation Sequencing, Sequencing, Transfection, Ubiquitin Assay, Over Expression

    8) Product Images from "MERS-CoV pathogenesis and antiviral efficacy of licensed drugs in human monocyte-derived antigen-presenting cells"

    Article Title: MERS-CoV pathogenesis and antiviral efficacy of licensed drugs in human monocyte-derived antigen-presenting cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0194868

    Antiviral activity of three compounds on MERS-CoV infection in MDMs and immature MDDCs. (A) Monocyte-derived macrophages (MDMs), (B) immature dendritic cells (MDDCs) and (C) Vero E6 cells were pretreated with toremifene (TOMF), chloroquine (CQ), and chlorpromazine (CPZ) for 1 h followed by inoculation with MERS-CoV Jordan variant at an MOI = 0.1. Supernatants collected at 72 h post infection (hpi) for APCs and 48 hpi for Vero E6 cells were inoculated onto Vero E6 cells pre-seeded overnight in 96 well plates and incubated for 6 days. Wells with cytopathic effects were counted, and TCID 50 was determined using the Reed-Muench method. Cytotoxicity was measured in parallel using luminescent cell viability assay at 48 h after compounds addition to the mock-infected MDMs and immature MDDCs. Antiviral activity is shown in gray and cytotoxicity is shown in orange. Results are representative of 2 individual experiments with 3 replicates in each (means ± standard error of the means [SEM]).
    Figure Legend Snippet: Antiviral activity of three compounds on MERS-CoV infection in MDMs and immature MDDCs. (A) Monocyte-derived macrophages (MDMs), (B) immature dendritic cells (MDDCs) and (C) Vero E6 cells were pretreated with toremifene (TOMF), chloroquine (CQ), and chlorpromazine (CPZ) for 1 h followed by inoculation with MERS-CoV Jordan variant at an MOI = 0.1. Supernatants collected at 72 h post infection (hpi) for APCs and 48 hpi for Vero E6 cells were inoculated onto Vero E6 cells pre-seeded overnight in 96 well plates and incubated for 6 days. Wells with cytopathic effects were counted, and TCID 50 was determined using the Reed-Muench method. Cytotoxicity was measured in parallel using luminescent cell viability assay at 48 h after compounds addition to the mock-infected MDMs and immature MDDCs. Antiviral activity is shown in gray and cytotoxicity is shown in orange. Results are representative of 2 individual experiments with 3 replicates in each (means ± standard error of the means [SEM]).

    Techniques Used: Activity Assay, Infection, Derivative Assay, Variant Assay, Incubation, Endpoint Dilution Assay, Cell Viability Assay

    9) Product Images from "The Human Sodium Iodide Symporter as a Reporter Gene for Studying Middle East Respiratory Syndrome Coronavirus Pathogenesis"

    Article Title: The Human Sodium Iodide Symporter as a Reporter Gene for Studying Middle East Respiratory Syndrome Coronavirus Pathogenesis

    Journal: mSphere

    doi: 10.1128/mSphere.00540-18

    Radio-uptake of 99m Tc-pertechnetate by planar scintigraphy. (a) Experimental overview of in vitro evaluation of the rMERS-CoV/ hNIS virus. Vero E6 cells were infected with rMERS-CoV or rMERS-CoV/ hNIS at an MOI of 0.01 or 0.04. At various time points postinfection, the cells were incubated with 99m Tc-pertechnetate, and images of the plates were acquired. (b) Plate layout for hNIS functional assays. (c) Representative images of the plates acquired at 24 h postinfection at an MOI of 0.01 (top plates) or 0.04 (bottom plates) after incubation with 99m Tc-pertechnetate.
    Figure Legend Snippet: Radio-uptake of 99m Tc-pertechnetate by planar scintigraphy. (a) Experimental overview of in vitro evaluation of the rMERS-CoV/ hNIS virus. Vero E6 cells were infected with rMERS-CoV or rMERS-CoV/ hNIS at an MOI of 0.01 or 0.04. At various time points postinfection, the cells were incubated with 99m Tc-pertechnetate, and images of the plates were acquired. (b) Plate layout for hNIS functional assays. (c) Representative images of the plates acquired at 24 h postinfection at an MOI of 0.01 (top plates) or 0.04 (bottom plates) after incubation with 99m Tc-pertechnetate.

    Techniques Used: In Vitro, Infection, Incubation, Functional Assay

    Retention of hNIS transgene following viral kinetics analysis and serial passage. (a and b) Vero E6 cells were infected with rMERS-CoV/hNIS (a) or parental rMERS-CoV (b) at an MOI of 0.01 or 3 and then collected at 96 h postinfection for RT-PCR. (c) Retention of the hNIS gene following serial passage. RNA was extracted from cells 72 h postinfection followed by RT-PCR at passage 6. A positive-control virus (C+) and uninfected negative-control cells (C−) were used as controls.
    Figure Legend Snippet: Retention of hNIS transgene following viral kinetics analysis and serial passage. (a and b) Vero E6 cells were infected with rMERS-CoV/hNIS (a) or parental rMERS-CoV (b) at an MOI of 0.01 or 3 and then collected at 96 h postinfection for RT-PCR. (c) Retention of the hNIS gene following serial passage. RNA was extracted from cells 72 h postinfection followed by RT-PCR at passage 6. A positive-control virus (C+) and uninfected negative-control cells (C−) were used as controls.

    Techniques Used: Infection, Reverse Transcription Polymerase Chain Reaction, Positive Control, Negative Control

    Kinetics of rMERS-CoV/ hNIS and parental rMERS-CoV replication in Vero E6 cells. (a and b) Multistep (a) and one-step (b) growth curves of Vero E6 cells infected with rMERS-CoV (Parental) and rMERS-CoV/ hNIS ( hNIS ). Quantification of the release of infectious virus at the indicated time points (hours postexposure) was determined by plaque assays. Each data point represents the mean ± standard deviation (SD) (error bar) averaged from three independent experiments. (c and d) Cytopathology of rMERS-CoV and rMERS-CoV/ hNIS in Vero E6 cells. The cells were infected with either rMERS-CoV or rMERS-CoV/ hNIS at an MOI of 0.01 (c) or 3 (d) and analyzed by light microscopy at the indicated time points. Photomicrographs were taken using a 40× objective.
    Figure Legend Snippet: Kinetics of rMERS-CoV/ hNIS and parental rMERS-CoV replication in Vero E6 cells. (a and b) Multistep (a) and one-step (b) growth curves of Vero E6 cells infected with rMERS-CoV (Parental) and rMERS-CoV/ hNIS ( hNIS ). Quantification of the release of infectious virus at the indicated time points (hours postexposure) was determined by plaque assays. Each data point represents the mean ± standard deviation (SD) (error bar) averaged from three independent experiments. (c and d) Cytopathology of rMERS-CoV and rMERS-CoV/ hNIS in Vero E6 cells. The cells were infected with either rMERS-CoV or rMERS-CoV/ hNIS at an MOI of 0.01 (c) or 3 (d) and analyzed by light microscopy at the indicated time points. Photomicrographs were taken using a 40× objective.

    Techniques Used: Infection, Standard Deviation, Light Microscopy

    10) Product Images from "Isolation and Identification of Rickettsia massiliae from Rhipicephalus sanguineus Ticks Collected in Arizona"

    Article Title: Isolation and Identification of Rickettsia massiliae from Rhipicephalus sanguineus Ticks Collected in Arizona

    Journal: Applied and Environmental Microbiology

    doi: 10.1128/AEM.00122-06

    Kinetics of AZT80 growth in XTC-2 cells (A) and VERO E6 cells (B) determined by the SYBR green PCR assay. Data are the averages (± standard error) of two independent measurements for each of two 33-mm dishes infected under the same conditions
    Figure Legend Snippet: Kinetics of AZT80 growth in XTC-2 cells (A) and VERO E6 cells (B) determined by the SYBR green PCR assay. Data are the averages (± standard error) of two independent measurements for each of two 33-mm dishes infected under the same conditions

    Techniques Used: SYBR Green Assay, Polymerase Chain Reaction, Infection

    11) Product Images from "Route of Infection Influences Zika Virus Shedding in a Guinea Pig Model"

    Article Title: Route of Infection Influences Zika Virus Shedding in a Guinea Pig Model

    Journal: Cells

    doi: 10.3390/cells8111437

    Viral shedding in guinea pigs inoculated subcutaneous (SQ) and intravaginal (VAG) with 1x10 6 PFU ZIKV MR766. Virus titer, determined by plaque assay on Vero E6 cells, detected in: ( A ) tears; ( B ) vaginal secretions. Inset shows the graph with the Y-axis shifted to show average titer in vaginal secretions through 21 dpi in VAG infected animals.
    Figure Legend Snippet: Viral shedding in guinea pigs inoculated subcutaneous (SQ) and intravaginal (VAG) with 1x10 6 PFU ZIKV MR766. Virus titer, determined by plaque assay on Vero E6 cells, detected in: ( A ) tears; ( B ) vaginal secretions. Inset shows the graph with the Y-axis shifted to show average titer in vaginal secretions through 21 dpi in VAG infected animals.

    Techniques Used: Plaque Assay, Infection

    12) Product Images from "Neuraminidase-Inhibiting Antibody Titers Correlate with Protection from Heterologous Influenza Virus Strains of the Same Neuraminidase Subtype"

    Article Title: Neuraminidase-Inhibiting Antibody Titers Correlate with Protection from Heterologous Influenza Virus Strains of the Same Neuraminidase Subtype

    Journal: Journal of Virology

    doi: 10.1128/JVI.01006-18

    Generation and characterization of influenza antigen-expressing VSV replicons. (A) Phylogenetic relationship of influenza A virus NA genes included in this study. Amino acid sequences were aligned using ClustalW and a phylogenetic tree constructed using the neighbor-joining method and 1,000 bootstrapped replications with observed amino acid differences as the distance on the aligned sequences using MEGA7. (B) Scheme of the genomes of the parental VSV and the recombinant VSV*ΔG replicons. VSV*ΔG lacks the gene encoding the VSV G protein and includes the eGFP gene in an additional transcription unit, depicted in gray. VSV*ΔG(X) encodes the respective heterologous influenza virus antigens, depicted in dark gray. (C) Vero E6 cells were surface biotin labeled with EZ-Link sulfo-NHS-biotin (Pierce) 12 h after infection with the respective VSV*ΔG replicons, followed by cell lysis and immunoprecipitation of 50 μg of cell extracts with homologous mouse antiserum. Proteins in the pellet fraction were separated on an SDS-PAGE gel, and surface biotin-labeled proteins were visualized using an avidin-HRP secondary antibody. (D) Protein bands from three independent replicates were quantified and normalized relative to NA PR8 for each blot and are shown as fold changes. Error bars represent the standard deviations of the means. Groups were compared using one-way ANOVA with a Tukey posttest, and statistical significance is indicated (*, P
    Figure Legend Snippet: Generation and characterization of influenza antigen-expressing VSV replicons. (A) Phylogenetic relationship of influenza A virus NA genes included in this study. Amino acid sequences were aligned using ClustalW and a phylogenetic tree constructed using the neighbor-joining method and 1,000 bootstrapped replications with observed amino acid differences as the distance on the aligned sequences using MEGA7. (B) Scheme of the genomes of the parental VSV and the recombinant VSV*ΔG replicons. VSV*ΔG lacks the gene encoding the VSV G protein and includes the eGFP gene in an additional transcription unit, depicted in gray. VSV*ΔG(X) encodes the respective heterologous influenza virus antigens, depicted in dark gray. (C) Vero E6 cells were surface biotin labeled with EZ-Link sulfo-NHS-biotin (Pierce) 12 h after infection with the respective VSV*ΔG replicons, followed by cell lysis and immunoprecipitation of 50 μg of cell extracts with homologous mouse antiserum. Proteins in the pellet fraction were separated on an SDS-PAGE gel, and surface biotin-labeled proteins were visualized using an avidin-HRP secondary antibody. (D) Protein bands from three independent replicates were quantified and normalized relative to NA PR8 for each blot and are shown as fold changes. Error bars represent the standard deviations of the means. Groups were compared using one-way ANOVA with a Tukey posttest, and statistical significance is indicated (*, P

    Techniques Used: Expressing, Construct, Recombinant, Labeling, Infection, Lysis, Immunoprecipitation, SDS Page, Avidin-Biotin Assay

    13) Product Images from "Abelson Kinase Inhibitors Are Potent Inhibitors of Severe Acute Respiratory Syndrome Coronavirus and Middle East Respiratory Syndrome Coronavirus Fusion"

    Article Title: Abelson Kinase Inhibitors Are Potent Inhibitors of Severe Acute Respiratory Syndrome Coronavirus and Middle East Respiratory Syndrome Coronavirus Fusion

    Journal: Journal of Virology

    doi: 10.1128/JVI.01429-16

    Specific knockdown of Abl2, and not Abl1, significantly inhibits MERS-CoV and SARS-CoV replication. (A and B) Specific knockdown of ABl1 (A) and Abl2 (B) was achieved in Vero E6 cells. Numbers under bands are the percentage of each knocked down protein
    Figure Legend Snippet: Specific knockdown of Abl2, and not Abl1, significantly inhibits MERS-CoV and SARS-CoV replication. (A and B) Specific knockdown of ABl1 (A) and Abl2 (B) was achieved in Vero E6 cells. Numbers under bands are the percentage of each knocked down protein

    Techniques Used:

    Imatinib does not interfere with trafficking of live MERS-CoV or SARS-CoV to early or late endosomes, respectively. Vero E6 cells were infected with live SARS-CoV (A) or MERS-CoV (B), with or without a 1-h 10 μM imatinib pretreatment. For imaging
    Figure Legend Snippet: Imatinib does not interfere with trafficking of live MERS-CoV or SARS-CoV to early or late endosomes, respectively. Vero E6 cells were infected with live SARS-CoV (A) or MERS-CoV (B), with or without a 1-h 10 μM imatinib pretreatment. For imaging

    Techniques Used: Infection, Imaging

    14) Product Images from "Treating Influenza and SARS-CoV-2 via mRNA-encoded Cas13a"

    Article Title: Treating Influenza and SARS-CoV-2 via mRNA-encoded Cas13a

    Journal: bioRxiv

    doi: 10.1101/2020.04.24.060418

    SARS-CoV-2 SARS-CoV-2 targeted guide selection and in vitro testing a) Guide selection process for SARS-CoV-2. 19 sequences of SARS-CoV-2 from the Wuhan region were aligned with the Toronto 2 SARS-CoV isolate sequence, and regions in ORF1ab and N with complete coverage were isolated. Guide target sequences were then tiled across the conserved region and checked for correct folding. b) Cas13a-lbu mRNA along with each of 8 guides against N, 1 guide against ORF1ab (R5.1), or NTCR were transfected in Vero-E6 cells overnight prior to infection with SARS-CoV-2 (MOI 0.1). At 60 hpi, crystal violet stain was used to assess CPE. Percent live cell area was then plotted. Error bars indicate SD. *** indicates p≤0.0001 (one-way ANOVA with multiple comparisons to NTCR). c) Cas13a-lbu mRNA along with either guide N3.2 alone or in the indicated combinations was transfected in Vero-E6 cells overnight. GFP and dCas13a-lbu mRNA along with guide N3.2 was used as controls to demonstrate catalytic guide activity. Cells were infected with SARS-CoV-2 (MOI 0.1), and at 60 hpi crystal violet stain was used to assess CPE. Percent live cell area was then plotted. Error bars indicate SD. ** indicates p=0.0082; *** indicates p≤0.005 (one-way ANOVA with multiple comparisons to NTCR).
    Figure Legend Snippet: SARS-CoV-2 SARS-CoV-2 targeted guide selection and in vitro testing a) Guide selection process for SARS-CoV-2. 19 sequences of SARS-CoV-2 from the Wuhan region were aligned with the Toronto 2 SARS-CoV isolate sequence, and regions in ORF1ab and N with complete coverage were isolated. Guide target sequences were then tiled across the conserved region and checked for correct folding. b) Cas13a-lbu mRNA along with each of 8 guides against N, 1 guide against ORF1ab (R5.1), or NTCR were transfected in Vero-E6 cells overnight prior to infection with SARS-CoV-2 (MOI 0.1). At 60 hpi, crystal violet stain was used to assess CPE. Percent live cell area was then plotted. Error bars indicate SD. *** indicates p≤0.0001 (one-way ANOVA with multiple comparisons to NTCR). c) Cas13a-lbu mRNA along with either guide N3.2 alone or in the indicated combinations was transfected in Vero-E6 cells overnight. GFP and dCas13a-lbu mRNA along with guide N3.2 was used as controls to demonstrate catalytic guide activity. Cells were infected with SARS-CoV-2 (MOI 0.1), and at 60 hpi crystal violet stain was used to assess CPE. Percent live cell area was then plotted. Error bars indicate SD. ** indicates p=0.0082; *** indicates p≤0.005 (one-way ANOVA with multiple comparisons to NTCR).

    Techniques Used: Selection, In Vitro, Sequencing, Isolation, Transfection, Infection, Staining, Activity Assay

    15) Product Images from "Cell attenuated porcine epidemic diarrhea virus strain Zhejiang08 provides effective immune protection attributed to dendritic cell stimulation"

    Article Title: Cell attenuated porcine epidemic diarrhea virus strain Zhejiang08 provides effective immune protection attributed to dendritic cell stimulation

    Journal: Vaccine

    doi: 10.1016/j.vaccine.2017.10.052

    Infectivity of Zhejang08 in Vero E6 cells. (A) CPE of Zhejiang08 in Vero E6 cells. Scale bars represent 50 μm. (B) Immunofluorescence images of PEDV (green) infected Vero E6 cells at 2, 12, 24, and 48 h post-infection (hpi). The scale bars represent 20 μm. (C) Viral plaque assay titer and M gene expression of PEDV infected Vero E6 cells (MOI = 0.1) at different infection times. Data express means ± SD (n = 3). Expression of the M gene was determined via quantitative RT-PCR using GAPDH gene expression for comparison. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
    Figure Legend Snippet: Infectivity of Zhejang08 in Vero E6 cells. (A) CPE of Zhejiang08 in Vero E6 cells. Scale bars represent 50 μm. (B) Immunofluorescence images of PEDV (green) infected Vero E6 cells at 2, 12, 24, and 48 h post-infection (hpi). The scale bars represent 20 μm. (C) Viral plaque assay titer and M gene expression of PEDV infected Vero E6 cells (MOI = 0.1) at different infection times. Data express means ± SD (n = 3). Expression of the M gene was determined via quantitative RT-PCR using GAPDH gene expression for comparison. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

    Techniques Used: Infection, Immunofluorescence, Viral Plaque Assay, Expressing, Quantitative RT-PCR

    16) Product Images from "Rational Design of Human Metapneumovirus Live Attenuated Vaccine Candidates by Inhibiting Viral mRNA Cap Methyltransferase"

    Article Title: Rational Design of Human Metapneumovirus Live Attenuated Vaccine Candidates by Inhibiting Viral mRNA Cap Methyltransferase

    Journal: Journal of Virology

    doi: 10.1128/JVI.00876-14

    Analysis of hMPV and VSV mRNA cap methylation by in vitro trans -methylation assay. (A) trans -G-N-7 methylation assay for hMPV and VSV. Five hundred nanograms of mRNA was isolated from virus-infected Vero E6 cells and was trans -methylated by vaccinia virus
    Figure Legend Snippet: Analysis of hMPV and VSV mRNA cap methylation by in vitro trans -methylation assay. (A) trans -G-N-7 methylation assay for hMPV and VSV. Five hundred nanograms of mRNA was isolated from virus-infected Vero E6 cells and was trans -methylated by vaccinia virus

    Techniques Used: Methylation, In Vitro, Isolation, Infection

    Sensitivity of recombinant hMPVs to IFN-α (A) and IFN-β (B) treatment. Confluent Vero E6 cells in 24-well plates were pretreated with DMEM containing 2 or 20 U of IFN-α or IFN-β at 37°C for 4 h. Cells were then
    Figure Legend Snippet: Sensitivity of recombinant hMPVs to IFN-α (A) and IFN-β (B) treatment. Confluent Vero E6 cells in 24-well plates were pretreated with DMEM containing 2 or 20 U of IFN-α or IFN-β at 37°C for 4 h. Cells were then

    Techniques Used: Recombinant

    17) Product Images from "Zika Virus Persistently Infects and Is Basolaterally Released from Primary Human Brain Microvascular Endothelial Cells"

    Article Title: Zika Virus Persistently Infects and Is Basolaterally Released from Primary Human Brain Microvascular Endothelial Cells

    Journal: mBio

    doi: 10.1128/mBio.00952-17

    hBMECs are viable after ZIKV infection. (A) Vero E6 or hBMECs were infected with ZIKV (MOI, 10), and costained 3 dpi with calcein-AM (green [live cells])/propidum iodide (red [dead cells]). Following calcein-AM/PI staining, monolayers were fixed and immunostained for ZIKV antigen. (B) Viability of ZIKV-infected Vero E6 cells and hBMECs was assessed via CyQuant NF uptake 3 dpi, and results were compared to those for mock-infected controls.
    Figure Legend Snippet: hBMECs are viable after ZIKV infection. (A) Vero E6 or hBMECs were infected with ZIKV (MOI, 10), and costained 3 dpi with calcein-AM (green [live cells])/propidum iodide (red [dead cells]). Following calcein-AM/PI staining, monolayers were fixed and immunostained for ZIKV antigen. (B) Viability of ZIKV-infected Vero E6 cells and hBMECs was assessed via CyQuant NF uptake 3 dpi, and results were compared to those for mock-infected controls.

    Techniques Used: Infection, Staining, CyQUANT Assay

    ZIKV persistently infects viable hBMECs 9 dpi. (A) hBMECs were infected with ZIKV as described for Fig. 2A , and titers present in cell supernatants were compared 2 to 9 dpi. (B and C) hBMECs were infected as described above, and cell lysates were analyzed for ZIKV RNA by qRT-PCR (B) and for ZIKV envelope protein (anti-Env) by Western blotting (C). Results were compared to those for the GAPDH controls 1 to 9 dpi. (D and E) Vero E6 cells or hBMECs were analyzed 9 dpi for ZIKV antigen and via calcein-AM/PI stain (D) or in CyQuant assays (E) for cell viability.
    Figure Legend Snippet: ZIKV persistently infects viable hBMECs 9 dpi. (A) hBMECs were infected with ZIKV as described for Fig. 2A , and titers present in cell supernatants were compared 2 to 9 dpi. (B and C) hBMECs were infected as described above, and cell lysates were analyzed for ZIKV RNA by qRT-PCR (B) and for ZIKV envelope protein (anti-Env) by Western blotting (C). Results were compared to those for the GAPDH controls 1 to 9 dpi. (D and E) Vero E6 cells or hBMECs were analyzed 9 dpi for ZIKV antigen and via calcein-AM/PI stain (D) or in CyQuant assays (E) for cell viability.

    Techniques Used: Infection, Quantitative RT-PCR, Western Blot, Staining, CyQUANT Assay

    ZIKV-infected hBMECs are viable and productive following cellular passage. (A and B) ZIKV-infected Vero E6 cells or hBMECs (MOI, 10) were trypsinized and passaged (1:3) 3 dpi and every 3 days thereafter. ZIKV-infected passaged hBMECs were detected by immunostaining. (C) Cells were infected and passaged as for panels A and B, and cell viability was assessed via calcein-AM/PI staining and fluorescent image overlay. (D) ZIKV titers in supernatants of hBMECs consecutively passaged 1 to 3 times (every 3 days).
    Figure Legend Snippet: ZIKV-infected hBMECs are viable and productive following cellular passage. (A and B) ZIKV-infected Vero E6 cells or hBMECs (MOI, 10) were trypsinized and passaged (1:3) 3 dpi and every 3 days thereafter. ZIKV-infected passaged hBMECs were detected by immunostaining. (C) Cells were infected and passaged as for panels A and B, and cell viability was assessed via calcein-AM/PI staining and fluorescent image overlay. (D) ZIKV titers in supernatants of hBMECs consecutively passaged 1 to 3 times (every 3 days).

    Techniques Used: Infection, Immunostaining, Staining

    Zika virus infection of primary hBMECs. (A) Primary hBMECs were infected with ZIKV (PRVABC59) at an MOI of 10, and 12 to 72 hpi ZIKV antigen-positive cells were detected by anti-DENV4 HMAF. (B to D) Titers of ZIKV-infected hBMEC supernatants were determined in an FFU assay (B) and analyzed for cellular ZIKV RNA levels by qRT-PCR (C) and for infected cells (D). (E to G) hBMECs and Vero E6 cells were pretreated with IFN-α (1,000 U/ml) for 3 h prior to ZIKV infection (MOI, 10) (E), or IFN-α was added at the indicated time postinfection and infected Vero E6 (F) or hBMECs (G) were immunostained and quantitated 24 later.
    Figure Legend Snippet: Zika virus infection of primary hBMECs. (A) Primary hBMECs were infected with ZIKV (PRVABC59) at an MOI of 10, and 12 to 72 hpi ZIKV antigen-positive cells were detected by anti-DENV4 HMAF. (B to D) Titers of ZIKV-infected hBMEC supernatants were determined in an FFU assay (B) and analyzed for cellular ZIKV RNA levels by qRT-PCR (C) and for infected cells (D). (E to G) hBMECs and Vero E6 cells were pretreated with IFN-α (1,000 U/ml) for 3 h prior to ZIKV infection (MOI, 10) (E), or IFN-α was added at the indicated time postinfection and infected Vero E6 (F) or hBMECs (G) were immunostained and quantitated 24 later.

    Techniques Used: Infection, Quantitative RT-PCR

    18) Product Images from "Assessing the Contributions of Inactivation, Removal, and Transfer of Ebola Virus and Vesicular Stomatitis Virus by Disinfectant Pre-soaked Wipes"

    Article Title: Assessing the Contributions of Inactivation, Removal, and Transfer of Ebola Virus and Vesicular Stomatitis Virus by Disinfectant Pre-soaked Wipes

    Journal: Frontiers in Public Health

    doi: 10.3389/fpubh.2020.00183

    Appearance of viral CPE or GFP in Vero E6 cells 2–14 days post-inoculation with EBOV/Mak.
    Figure Legend Snippet: Appearance of viral CPE or GFP in Vero E6 cells 2–14 days post-inoculation with EBOV/Mak.

    Techniques Used:

    19) Product Images from "Severe Acute Respiratory Syndrome-Related Coronavirus Is Inhibited by Interferon-α"

    Article Title: Severe Acute Respiratory Syndrome-Related Coronavirus Is Inhibited by Interferon-α

    Journal: The Journal of Infectious Diseases

    doi: 10.1086/382597

    Effect of interferon (IFN)-α-2b on virus growth. A , Reduction of virus yield in the presence of antiviral activity. Vero E6 cells were infected with ∼100 pfu of the severe acute respiratory syndrome-related coronavirus (SARS CoV). At 1 h after infection, the supernatants were replaced with Dulbecco's modified Eagle's medium (DMEM) or DMEM containing the indicated amounts of IFN-α-2b (IU/mL) for a total of 72 h. Growth of the SARS CoV isolate Tor2 was determined by plaquetitration assay from 3 independent experiments, and data are mean ± SD. B , Analysis of cell-to-cell spread of SARS CoV in the presence and absence of IFN-α-2b. Vero E6 cell monolayers were infected with 10-fold dilutions of the SARS CoV. After 1 h of virus adsorption, the inoculum was removed, and cell monolayers were overlaid with 0.9% low-meltingpoint agarose in DMEM (B2) or DMEM containing 5000 IU/mL of IFNa- 2b (B1). On day 3 after infection, cells were stained with crystal violet. C , Protein analysis. Cell lysates were subjected to 10% SDS-PAGE. Protein was electrotransferred to polyvinylidene difluoride membrane (Immobilon P membrane; Millipore). Viral antigen was detected by use of a patient serum sample and horseradish peroxidase-conjugated anti-human IgG, by use of enhanced chemiluminescence. NP, nucleoprotein. D , Comparison of the effect of antiviral activity on different SARS CoV isolates. Vero E6 cells were infected with 4 independently isolated SARS CoVs (isolates Tor2, Tor3, Tor7, and Tor684) and were analyzed for their sensitivity to IFN-α-2b, as described above.
    Figure Legend Snippet: Effect of interferon (IFN)-α-2b on virus growth. A , Reduction of virus yield in the presence of antiviral activity. Vero E6 cells were infected with ∼100 pfu of the severe acute respiratory syndrome-related coronavirus (SARS CoV). At 1 h after infection, the supernatants were replaced with Dulbecco's modified Eagle's medium (DMEM) or DMEM containing the indicated amounts of IFN-α-2b (IU/mL) for a total of 72 h. Growth of the SARS CoV isolate Tor2 was determined by plaquetitration assay from 3 independent experiments, and data are mean ± SD. B , Analysis of cell-to-cell spread of SARS CoV in the presence and absence of IFN-α-2b. Vero E6 cell monolayers were infected with 10-fold dilutions of the SARS CoV. After 1 h of virus adsorption, the inoculum was removed, and cell monolayers were overlaid with 0.9% low-meltingpoint agarose in DMEM (B2) or DMEM containing 5000 IU/mL of IFNa- 2b (B1). On day 3 after infection, cells were stained with crystal violet. C , Protein analysis. Cell lysates were subjected to 10% SDS-PAGE. Protein was electrotransferred to polyvinylidene difluoride membrane (Immobilon P membrane; Millipore). Viral antigen was detected by use of a patient serum sample and horseradish peroxidase-conjugated anti-human IgG, by use of enhanced chemiluminescence. NP, nucleoprotein. D , Comparison of the effect of antiviral activity on different SARS CoV isolates. Vero E6 cells were infected with 4 independently isolated SARS CoVs (isolates Tor2, Tor3, Tor7, and Tor684) and were analyzed for their sensitivity to IFN-α-2b, as described above.

    Techniques Used: Activity Assay, Infection, Modification, Adsorption, Staining, SDS Page, Isolation

    Cytopathic effect in severe acute respiratory syndrome-related coronavirus (SARS CoV)-infected Vero E6 cells. Vero E6 cells were infected with the SARS CoV at an MOI of 0.001 for 1 h and then were incubated in Dulbecco's modified Eagle's medium containing the indicated amounts of interferon-α-2b (IU/mL). Cells were fixed with 4% formalin on day 3 after infection.
    Figure Legend Snippet: Cytopathic effect in severe acute respiratory syndrome-related coronavirus (SARS CoV)-infected Vero E6 cells. Vero E6 cells were infected with the SARS CoV at an MOI of 0.001 for 1 h and then were incubated in Dulbecco's modified Eagle's medium containing the indicated amounts of interferon-α-2b (IU/mL). Cells were fixed with 4% formalin on day 3 after infection.

    Techniques Used: Infection, Incubation, Modification

    20) Product Images from "Severe Acute Respiratory Syndrome Coronavirus ORF7a Inhibits Bone Marrow Stromal Antigen 2 Virion Tethering through a Novel Mechanism of Glycosylation Interference"

    Article Title: Severe Acute Respiratory Syndrome Coronavirus ORF7a Inhibits Bone Marrow Stromal Antigen 2 Virion Tethering through a Novel Mechanism of Glycosylation Interference

    Journal: Journal of Virology

    doi: 10.1128/JVI.02274-15

    BST-2 tethers SARS-CoV to the plasma membrane. BST-2 or control plasmid was transfected into Vero E6 cells and infected with either icSARS-CoV or icSARS-ORF7abΔ-CoV at an MOI of 10. At 24 h postinfection, cells were fixed and imaged by electron
    Figure Legend Snippet: BST-2 tethers SARS-CoV to the plasma membrane. BST-2 or control plasmid was transfected into Vero E6 cells and infected with either icSARS-CoV or icSARS-ORF7abΔ-CoV at an MOI of 10. At 24 h postinfection, cells were fixed and imaged by electron

    Techniques Used: Plasmid Preparation, Transfection, Infection

    21) Product Images from "Involvement of Surfactant Protein D in Ebola Virus Infection Enhancement via Glycoprotein Interaction"

    Article Title: Involvement of Surfactant Protein D in Ebola Virus Infection Enhancement via Glycoprotein Interaction

    Journal: Viruses

    doi: 10.3390/v11010015

    Transduction of non-replicative GP-R pseudoparticles in presence of hSP-D and pSP-D. rVSV-RFP-GP-R pseudoparticles were preincubated with hSP-D and pSP-D (10µg/mL) for 1 h at 37 °C before incubation with Vero E6 cells ( A ) or A549 cells ( B ) for 1 h at 37 °C. RFP level expression was analyzed using flow cytometry. The results for the preincubated groups were compared to those of non-preincubated groups. *, a statistically difference ( p
    Figure Legend Snippet: Transduction of non-replicative GP-R pseudoparticles in presence of hSP-D and pSP-D. rVSV-RFP-GP-R pseudoparticles were preincubated with hSP-D and pSP-D (10µg/mL) for 1 h at 37 °C before incubation with Vero E6 cells ( A ) or A549 cells ( B ) for 1 h at 37 °C. RFP level expression was analyzed using flow cytometry. The results for the preincubated groups were compared to those of non-preincubated groups. *, a statistically difference ( p

    Techniques Used: Transduction, Incubation, Expressing, Flow Cytometry, Cytometry

    Enhancement of replicative-GP virus infection. wtVSV-G, rVSV-GP, and EBOV were preincubated with collectins for 1 h at 37 °C before the infection of Vero E6 cells for 1 h at 37 °C at a MOI of 5 × 10 −4 , 1 × 10 −4 , and 2 x 10 5 in 24-well culture plates, respectively. Cells were infected with rVSV-GP preincubated with ( A ) AP-SP-A, hSP-D, and MBL (10 µg/mL), ( B ) increasing concentrations of hSP-D (5, 10, and 20 µg/mL), or ( C ) pSP-D (10 µg/mL). (A, B, and C) At 2 days post-infection, rVSV-GP replication was measured through determination of the PFU number. ( D ) Cells were infected with wt-EBOV preincubated with increasing concentrations of hSP-D (2.5, 5, and 10 µg/mL), hNCRD (100µg/mL and 300µg/mL), and AP-SP-A (10 µg/mL). ( E ) Cells were infected with purified-EBOV preincubated with increasing concentrations of AP-SP-A, hSP-D, and pSP-D (1, 5, and 10 µg/mL). EBOV replication was measured at 6 days post-infection using an IHC assay. The results for the preincubated groups were compared to those for the nonpreincubated groups. *, a statistically difference ( p
    Figure Legend Snippet: Enhancement of replicative-GP virus infection. wtVSV-G, rVSV-GP, and EBOV were preincubated with collectins for 1 h at 37 °C before the infection of Vero E6 cells for 1 h at 37 °C at a MOI of 5 × 10 −4 , 1 × 10 −4 , and 2 x 10 5 in 24-well culture plates, respectively. Cells were infected with rVSV-GP preincubated with ( A ) AP-SP-A, hSP-D, and MBL (10 µg/mL), ( B ) increasing concentrations of hSP-D (5, 10, and 20 µg/mL), or ( C ) pSP-D (10 µg/mL). (A, B, and C) At 2 days post-infection, rVSV-GP replication was measured through determination of the PFU number. ( D ) Cells were infected with wt-EBOV preincubated with increasing concentrations of hSP-D (2.5, 5, and 10 µg/mL), hNCRD (100µg/mL and 300µg/mL), and AP-SP-A (10 µg/mL). ( E ) Cells were infected with purified-EBOV preincubated with increasing concentrations of AP-SP-A, hSP-D, and pSP-D (1, 5, and 10 µg/mL). EBOV replication was measured at 6 days post-infection using an IHC assay. The results for the preincubated groups were compared to those for the nonpreincubated groups. *, a statistically difference ( p

    Techniques Used: Infection, Purification, Immunohistochemistry

    22) Product Images from "Zinc Binding Activity of Human Metapneumovirus M2-1 Protein Is Indispensable for Viral Replication and Pathogenesis In Vivo"

    Article Title: Zinc Binding Activity of Human Metapneumovirus M2-1 Protein Is Indispensable for Viral Replication and Pathogenesis In Vivo

    Journal: Journal of Virology

    doi: 10.1128/JVI.03488-14

    Multistep growth curve of recombinant hMPVs carrying mutations in the zinc binding site. Vero E6 cells in 35-mm dishes were infected with each recombinant hMPV at an MOI of 0.01. After adsorption for 1 h, the inoculums were removed and the infected cells
    Figure Legend Snippet: Multistep growth curve of recombinant hMPVs carrying mutations in the zinc binding site. Vero E6 cells in 35-mm dishes were infected with each recombinant hMPV at an MOI of 0.01. After adsorption for 1 h, the inoculums were removed and the infected cells

    Techniques Used: Recombinant, Binding Assay, Infection, Adsorption

    23) Product Images from "Neutrophil Activation in Acute Hemorrhagic Fever With Renal Syndrome Is Mediated by Hantavirus-Infected Microvascular Endothelial Cells"

    Article Title: Neutrophil Activation in Acute Hemorrhagic Fever With Renal Syndrome Is Mediated by Hantavirus-Infected Microvascular Endothelial Cells

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2018.02098

    Antiviral activity of activated PMNs. Purified PUUV was incubated with quiescent or PMA-activated freshly isolated PMNs in the presence of inhibitors for MPO (0.01 or 0.002% NaN 3 ), serine proteases (1 or 0.2 mM PMSF), metalloproteinases (1 mM EDTA) or NETs (10 U/ml DNAse) and (A) subjected to immunoblotting for viral proteins Gn, Gc and N protein and infectious titer measurement in Vero E6 cells (B) . C, Control without the addition of PMNs.
    Figure Legend Snippet: Antiviral activity of activated PMNs. Purified PUUV was incubated with quiescent or PMA-activated freshly isolated PMNs in the presence of inhibitors for MPO (0.01 or 0.002% NaN 3 ), serine proteases (1 or 0.2 mM PMSF), metalloproteinases (1 mM EDTA) or NETs (10 U/ml DNAse) and (A) subjected to immunoblotting for viral proteins Gn, Gc and N protein and infectious titer measurement in Vero E6 cells (B) . C, Control without the addition of PMNs.

    Techniques Used: Activity Assay, Purification, Incubation, Isolation

    24) Product Images from "Methyltransferase-Defective Avian Metapneumovirus Vaccines Provide Complete Protection against Challenge with the Homologous Colorado Strain and the Heterologous Minnesota Strain"

    Article Title: Methyltransferase-Defective Avian Metapneumovirus Vaccines Provide Complete Protection against Challenge with the Homologous Colorado Strain and the Heterologous Minnesota Strain

    Journal: Journal of Virology

    doi: 10.1128/JVI.01095-14

    Plaque morphology of recombinant aMPVs carrying mutations in the SAM binding site. An agarose overlay plaque assay was performed in monolayer Vero-E6 cells. Viral plaques were developed at day 7 postinfection. The cells were fixed in 10% formaldehyde,
    Figure Legend Snippet: Plaque morphology of recombinant aMPVs carrying mutations in the SAM binding site. An agarose overlay plaque assay was performed in monolayer Vero-E6 cells. Viral plaques were developed at day 7 postinfection. The cells were fixed in 10% formaldehyde,

    Techniques Used: Recombinant, Binding Assay, Plaque Assay

    Multistep growth curve of recombinant hMPVs carrying mutations in the SAM binding site. Vero-E6 cells in 35-mm dishes were infected with each recombinant raMPV at an MOI of 0.1. After adsorption for 1 h, the inocula were removed, and the infected cells
    Figure Legend Snippet: Multistep growth curve of recombinant hMPVs carrying mutations in the SAM binding site. Vero-E6 cells in 35-mm dishes were infected with each recombinant raMPV at an MOI of 0.1. After adsorption for 1 h, the inocula were removed, and the infected cells

    Techniques Used: Recombinant, Binding Assay, Infection, Adsorption

    Cytopathic effects (CPEs) caused by raMPVs carrying mutations in the SAM binding site. Vero-E6 cells were infected with each recombinant aMPV at an MOI of 0.1. CPE was monitored on a daily basis. Pictures were taken at days 2, 4, and 6 postinfection.
    Figure Legend Snippet: Cytopathic effects (CPEs) caused by raMPVs carrying mutations in the SAM binding site. Vero-E6 cells were infected with each recombinant aMPV at an MOI of 0.1. CPE was monitored on a daily basis. Pictures were taken at days 2, 4, and 6 postinfection.

    Techniques Used: Binding Assay, Infection, Recombinant

    25) Product Images from "Isolation and partial characterization of a highly divergent lineage of hantavirus from the European mole (Talpa europaea)"

    Article Title: Isolation and partial characterization of a highly divergent lineage of hantavirus from the European mole (Talpa europaea)

    Journal: Scientific Reports

    doi: 10.1038/srep21119

    ( A ) European mole ( Talpa europaea ) captured in Huta Dłutowska, in central Poland, in August 2013. ( B ) Thin-section transmission electron micrograph, showing hantavirus particles in Vero E6 cells inoculated with lung homogenate from a NVAV-infected European mole. Bar = 100 nm.
    Figure Legend Snippet: ( A ) European mole ( Talpa europaea ) captured in Huta Dłutowska, in central Poland, in August 2013. ( B ) Thin-section transmission electron micrograph, showing hantavirus particles in Vero E6 cells inoculated with lung homogenate from a NVAV-infected European mole. Bar = 100 nm.

    Techniques Used: Transmission Assay, Infection

    26) Product Images from "Neutralization of genotype 2 porcine epidemic diarrhea virus strains by a novel monoclonal antibody"

    Article Title: Neutralization of genotype 2 porcine epidemic diarrhea virus strains by a novel monoclonal antibody

    Journal: Virology

    doi: 10.1016/j.virol.2017.04.026

    Reaction of mAb 1B9 with PEDV-positive cells, as detected by an IFA. The cells in A) and C) were Vero E6 cells infected by PEDV strain LNCT2, which yielded positive signals; the cells in B) and D) were healthy Vero E6 cells used as a negative control. In A) and B), mAb 1B9 was used as the primary antibody, while in C) and D), a PEDV polyclonal antibody was used as the primary antibody.
    Figure Legend Snippet: Reaction of mAb 1B9 with PEDV-positive cells, as detected by an IFA. The cells in A) and C) were Vero E6 cells infected by PEDV strain LNCT2, which yielded positive signals; the cells in B) and D) were healthy Vero E6 cells used as a negative control. In A) and B), mAb 1B9 was used as the primary antibody, while in C) and D), a PEDV polyclonal antibody was used as the primary antibody.

    Techniques Used: Immunofluorescence, Infection, Negative Control

    27) Product Images from "Tropism and replication of Middle East respiratory syndrome coronavirus from dromedary camels in the human respiratory tract: an in-vitro and ex-vivo study"

    Article Title: Tropism and replication of Middle East respiratory syndrome coronavirus from dromedary camels in the human respiratory tract: an in-vitro and ex-vivo study

    Journal: The Lancet. Respiratory Medicine

    doi: 10.1016/S2213-2600(14)70158-4

    Replication kinetics of the human and camel MERS-CoV strains in Vero-E6 cells Vero-E6 cells were infected with the indicated viruses at multiplicity of infections of 0·01 and 2 and cultured at 37°C for 72 h. Culture supernatants were harvested at the indicated times, and virus titres were measured by TCID 50 assay. Results are presented as individual replicate (mean [SEM] for the three experiments). MOI=multiplicity of infection. TCID 50 =50% tissue culture infective dose.
    Figure Legend Snippet: Replication kinetics of the human and camel MERS-CoV strains in Vero-E6 cells Vero-E6 cells were infected with the indicated viruses at multiplicity of infections of 0·01 and 2 and cultured at 37°C for 72 h. Culture supernatants were harvested at the indicated times, and virus titres were measured by TCID 50 assay. Results are presented as individual replicate (mean [SEM] for the three experiments). MOI=multiplicity of infection. TCID 50 =50% tissue culture infective dose.

    Techniques Used: Infection, Cell Culture

    28) Product Images from "Gastrointestinal Tract As Entry Route for Hantavirus Infection"

    Article Title: Gastrointestinal Tract As Entry Route for Hantavirus Infection

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2017.01721

    Hantavirus survival in gastric juice. Infectious dose of 10 6 Puumala virus particles was suspended to pure gastric juice set to pH 1–7 with NaOH for 1, 10, or 15 min. After given incubation intervals the gastric juice was neutralized with NaOH and the virus suspension was used to infect VERO-E6 cells for focus titration of infectious particles ( n = 4). As at low pH the inactivation activity was expected to be high pH 2 was omitted to save valuable sample volume.
    Figure Legend Snippet: Hantavirus survival in gastric juice. Infectious dose of 10 6 Puumala virus particles was suspended to pure gastric juice set to pH 1–7 with NaOH for 1, 10, or 15 min. After given incubation intervals the gastric juice was neutralized with NaOH and the virus suspension was used to infect VERO-E6 cells for focus titration of infectious particles ( n = 4). As at low pH the inactivation activity was expected to be high pH 2 was omitted to save valuable sample volume.

    Techniques Used: Incubation, Titration, Activity Assay

    29) Product Images from "Processing of the Ebola virus glycoprotein by the proprotein convertase furin"

    Article Title: Processing of the Ebola virus glycoprotein by the proprotein convertase furin

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi:

    Virion GP of EBOV consists of two cleavage subunits. EBOV GP metabolically labeled with [ 3 H]glucosamine or with [ 35 S]methionine-cysteine was immunoprecipitated from purified virions grown in Vero E6 cells and analyzed by 15% SDS/PAGE under reducing or nonreducing conditions. The positions of the mature GP (GP 1,2 ), and the two cleavage subunits GP 1 and GP 2 , are indicated.
    Figure Legend Snippet: Virion GP of EBOV consists of two cleavage subunits. EBOV GP metabolically labeled with [ 3 H]glucosamine or with [ 35 S]methionine-cysteine was immunoprecipitated from purified virions grown in Vero E6 cells and analyzed by 15% SDS/PAGE under reducing or nonreducing conditions. The positions of the mature GP (GP 1,2 ), and the two cleavage subunits GP 1 and GP 2 , are indicated.

    Techniques Used: Metabolic Labelling, Labeling, Immunoprecipitation, Purification, SDS Page

    30) Product Images from "Multimeric single-domain antibody complexes protect against bunyavirus infections"

    Article Title: Multimeric single-domain antibody complexes protect against bunyavirus infections

    Journal: eLife

    doi: 10.7554/eLife.52716

    VHH specificity determined by immunoperoxidase monolayer assay (IPMA). Vero E6 cells were infected at MOI 0.1 with either RVFV Clone 13 (blue boxes) or SBV NL-F6 (green boxes) and fixed 1 day post infection following permeabilization and VHH incubation at 10 µg/ml. VHH binding was visualized with a HRP conjugated goat anti-llama IgG conjugate and a AEC (3-amino-9-ethylcarbazole) substrate.
    Figure Legend Snippet: VHH specificity determined by immunoperoxidase monolayer assay (IPMA). Vero E6 cells were infected at MOI 0.1 with either RVFV Clone 13 (blue boxes) or SBV NL-F6 (green boxes) and fixed 1 day post infection following permeabilization and VHH incubation at 10 µg/ml. VHH binding was visualized with a HRP conjugated goat anti-llama IgG conjugate and a AEC (3-amino-9-ethylcarbazole) substrate.

    Techniques Used: Infection, Incubation, Binding Assay

    31) Product Images from "Neutralizing Monoclonal Antibodies against the Gn and the Gc of the Andes Virus Glycoprotein Spike Complex Protect from Virus Challenge in a Preclinical Hamster Model"

    Article Title: Neutralizing Monoclonal Antibodies against the Gn and the Gc of the Andes Virus Glycoprotein Spike Complex Protect from Virus Challenge in a Preclinical Hamster Model

    Journal: mBio

    doi: 10.1128/mBio.00028-20

    Neutralization and effector functions of the isolated MAbs. (A) FRNAs of “AN” fusion MAbs against VSV-ANDV. (B) FRNAs of “HAP” fusion MAbs against VSV-ANDV. For both A and B, each MAb was run in duplicates in 3-fold serial dilutions starting at 30 μg/ml. Any MAb exhibiting > 15% neutralization at the last concentration was repeated with lower dilutions. The negative-control MAb was a murine IgG2a specific for Zaire ebolavirus (KL-2G12). All trend lines are logarithmic regressions except where such regressions were not converged. (C) FRNAs conducted as for panels A and B but against authentic ANDV. Dashed lines in panels A, B, and C demonstrate 50% inhibition. (D) Comparison of IC 50 values of each MAb against VSV-ANDV and authentic ANDV. Error bars indicate 95% confidence intervals. (E and F) ADCC reporter assays of each MAb against VSV-ANDV-infected Vero.E6 cells (MOI, 1.0). Data shown are the result from one experiment with a shared positive control in panels E and F (serum from homologous fusion). This positive control was used in 3-fold serial dilutions with a starting dilution of 1:300.
    Figure Legend Snippet: Neutralization and effector functions of the isolated MAbs. (A) FRNAs of “AN” fusion MAbs against VSV-ANDV. (B) FRNAs of “HAP” fusion MAbs against VSV-ANDV. For both A and B, each MAb was run in duplicates in 3-fold serial dilutions starting at 30 μg/ml. Any MAb exhibiting > 15% neutralization at the last concentration was repeated with lower dilutions. The negative-control MAb was a murine IgG2a specific for Zaire ebolavirus (KL-2G12). All trend lines are logarithmic regressions except where such regressions were not converged. (C) FRNAs conducted as for panels A and B but against authentic ANDV. Dashed lines in panels A, B, and C demonstrate 50% inhibition. (D) Comparison of IC 50 values of each MAb against VSV-ANDV and authentic ANDV. Error bars indicate 95% confidence intervals. (E and F) ADCC reporter assays of each MAb against VSV-ANDV-infected Vero.E6 cells (MOI, 1.0). Data shown are the result from one experiment with a shared positive control in panels E and F (serum from homologous fusion). This positive control was used in 3-fold serial dilutions with a starting dilution of 1:300.

    Techniques Used: Neutralization, Isolation, Concentration Assay, Negative Control, Inhibition, Infection, Positive Control

    32) Product Images from "Recombinant Lassa Virus Expressing Green Fluorescent Protein as a Tool for High-Throughput Drug Screens and Neutralizing Antibody Assays"

    Article Title: Recombinant Lassa Virus Expressing Green Fluorescent Protein as a Tool for High-Throughput Drug Screens and Neutralizing Antibody Assays

    Journal: Viruses

    doi: 10.3390/v10110655

    Stability of the GFP reporter gene expressed by rLASV-GFP. Cells were exposed to rLASV-GFP (MOI = 0.01). At 72 h PI, TCS were collected (passage 1, P1), and virus titers were determined by plaque assay. Fresh Vero ( a ) or Vero-E6 cells ( b ) were exposed to with TCS from P1 (MOI = 0.01). This process was serially repeated throughout P10. GFP expression (green) was measured by high-content imaging. Nuclei were stained with DAPI (blue). Viral RNA was extracted from TCS of P1 to P10 to amplify DNA fragments containing an GFP-P2A and a part of the NP ORF using RT-PCR. Images are representative field images of individual wells. Bar, 200 µm.
    Figure Legend Snippet: Stability of the GFP reporter gene expressed by rLASV-GFP. Cells were exposed to rLASV-GFP (MOI = 0.01). At 72 h PI, TCS were collected (passage 1, P1), and virus titers were determined by plaque assay. Fresh Vero ( a ) or Vero-E6 cells ( b ) were exposed to with TCS from P1 (MOI = 0.01). This process was serially repeated throughout P10. GFP expression (green) was measured by high-content imaging. Nuclei were stained with DAPI (blue). Viral RNA was extracted from TCS of P1 to P10 to amplify DNA fragments containing an GFP-P2A and a part of the NP ORF using RT-PCR. Images are representative field images of individual wells. Bar, 200 µm.

    Techniques Used: Plaque Assay, Expressing, Imaging, Staining, Reverse Transcription Polymerase Chain Reaction

    Antiviral drug evaluation based on recombinant LASV expressing GFP (rLASV-GFP). ( a ) Vero E6 cells were pretreated with drugs at the indicated concentrations and then exposed to rLASV-WT or rLASV-GFP (MOI = 0.1) in the presence of the drugs. Viral titers in TCS at 48 h PI were determined by plaque assay. Values represent the means ± standard deviations of triplicate samples. ( b ) Infectivity of rLASV-GFP in A549, HeLa, Huh7, and Vero E6 cells at the indicated MOIs at 24, 48, and 72h PI as determined by GFP-expression using high-content imaging. ( c ) Effect of favipiravir and ribavirin on rLASV-GFP multiplication at 48 (orange filled circles) and 72 h PI (green filled squares). Cells were exposed to rLASV-GFP (MOI = 0.1) and treated with various concentrations of favipiravir or ribavirin. The percentage of GFP-positive cells was determined at 48 h or 72 h PI. ( d ) Half-maximal effective concentrations (EC 50 ) of favipiravir and ribavirin to inhibit rLASV-GFP infection in four cell types at 48 and 72 h PI.
    Figure Legend Snippet: Antiviral drug evaluation based on recombinant LASV expressing GFP (rLASV-GFP). ( a ) Vero E6 cells were pretreated with drugs at the indicated concentrations and then exposed to rLASV-WT or rLASV-GFP (MOI = 0.1) in the presence of the drugs. Viral titers in TCS at 48 h PI were determined by plaque assay. Values represent the means ± standard deviations of triplicate samples. ( b ) Infectivity of rLASV-GFP in A549, HeLa, Huh7, and Vero E6 cells at the indicated MOIs at 24, 48, and 72h PI as determined by GFP-expression using high-content imaging. ( c ) Effect of favipiravir and ribavirin on rLASV-GFP multiplication at 48 (orange filled circles) and 72 h PI (green filled squares). Cells were exposed to rLASV-GFP (MOI = 0.1) and treated with various concentrations of favipiravir or ribavirin. The percentage of GFP-positive cells was determined at 48 h or 72 h PI. ( d ) Half-maximal effective concentrations (EC 50 ) of favipiravir and ribavirin to inhibit rLASV-GFP infection in four cell types at 48 and 72 h PI.

    Techniques Used: Recombinant, Expressing, Plaque Assay, Infection, Imaging

    33) Product Images from "Topographic Changes in SARS Coronavirus-infected Cells at Late Stages of Infection"

    Article Title: Topographic Changes in SARS Coronavirus-infected Cells at Late Stages of Infection

    Journal: Emerging Infectious Diseases

    doi: 10.3201/eid1011.040195

    Scanning electron microscopy of Vero E6 cells infected with severe acute respiratory syndrome–associated coronavirus at 15 h after infection. A) One pronounced surface morphologic change is the proliferation of psuedopodia at the cell periphery (arrows). Some pseudopodia are also developing on the cell surface. Some cells appear to have large amount of extracellular virus on the cell surface (arrowhead), whereas neighboring cells seem deprived of any extracelluar virus particles. B) Virus particles are protruding from the edge of cells (arrows). Inset shows the boxed area at higher magnification. Virus particles appear knobby and rosettelike.
    Figure Legend Snippet: Scanning electron microscopy of Vero E6 cells infected with severe acute respiratory syndrome–associated coronavirus at 15 h after infection. A) One pronounced surface morphologic change is the proliferation of psuedopodia at the cell periphery (arrows). Some pseudopodia are also developing on the cell surface. Some cells appear to have large amount of extracellular virus on the cell surface (arrowhead), whereas neighboring cells seem deprived of any extracelluar virus particles. B) Virus particles are protruding from the edge of cells (arrows). Inset shows the boxed area at higher magnification. Virus particles appear knobby and rosettelike.

    Techniques Used: Electron Microscopy, Infection

    Scanning electron microscopy of Vero E6 cells infected with severe acute respiratory syndrome–associated coronavirus at 24 h after infection. A) Cell surface is covered with extracellular progeny virus particles, and progeny virus are being extruded from or attached to numerous pseudopodia on infected cell surface (arrows). B) A higher magnification micrograph of the virus-clustered pseudopodia (arrows). C) Rosettelike appearance of the matured virus particles (arrows). The scanning electron microscopy image complements the form and structure of the virus seen with negative staining (inset) under transmission electron microscopy. Short and stubby spikes are visible on the virus surface. D) Arrows indicate virus particles being exported from the surfaces of the filopodia.
    Figure Legend Snippet: Scanning electron microscopy of Vero E6 cells infected with severe acute respiratory syndrome–associated coronavirus at 24 h after infection. A) Cell surface is covered with extracellular progeny virus particles, and progeny virus are being extruded from or attached to numerous pseudopodia on infected cell surface (arrows). B) A higher magnification micrograph of the virus-clustered pseudopodia (arrows). C) Rosettelike appearance of the matured virus particles (arrows). The scanning electron microscopy image complements the form and structure of the virus seen with negative staining (inset) under transmission electron microscopy. Short and stubby spikes are visible on the virus surface. D) Arrows indicate virus particles being exported from the surfaces of the filopodia.

    Techniques Used: Electron Microscopy, Infection, Negative Staining, Transmission Assay

    34) Product Images from "Neutrophil Activation in Acute Hemorrhagic Fever With Renal Syndrome Is Mediated by Hantavirus-Infected Microvascular Endothelial Cells"

    Article Title: Neutrophil Activation in Acute Hemorrhagic Fever With Renal Syndrome Is Mediated by Hantavirus-Infected Microvascular Endothelial Cells

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2018.02098

    Antiviral activity of activated PMNs. Purified PUUV was incubated with quiescent or PMA-activated freshly isolated PMNs in the presence of inhibitors for MPO (0.01 or 0.002% NaN 3 ), serine proteases (1 or 0.2 mM PMSF), metalloproteinases (1 mM EDTA) or NETs (10 U/ml DNAse) and (A) subjected to immunoblotting for viral proteins Gn, Gc and N protein and infectious titer measurement in Vero E6 cells (B) . C, Control without the addition of PMNs.
    Figure Legend Snippet: Antiviral activity of activated PMNs. Purified PUUV was incubated with quiescent or PMA-activated freshly isolated PMNs in the presence of inhibitors for MPO (0.01 or 0.002% NaN 3 ), serine proteases (1 or 0.2 mM PMSF), metalloproteinases (1 mM EDTA) or NETs (10 U/ml DNAse) and (A) subjected to immunoblotting for viral proteins Gn, Gc and N protein and infectious titer measurement in Vero E6 cells (B) . C, Control without the addition of PMNs.

    Techniques Used: Activity Assay, Purification, Incubation, Isolation

    35) Product Images from "The Human Sodium Iodide Symporter as a Reporter Gene for Studying Middle East Respiratory Syndrome Coronavirus Pathogenesis"

    Article Title: The Human Sodium Iodide Symporter as a Reporter Gene for Studying Middle East Respiratory Syndrome Coronavirus Pathogenesis

    Journal: mSphere

    doi: 10.1128/mSphere.00540-18

    Radio-uptake of 99m Tc-pertechnetate by planar scintigraphy. (a) Experimental overview of in vitro evaluation of the rMERS-CoV/ hNIS virus. Vero E6 cells were infected with rMERS-CoV or rMERS-CoV/ hNIS at an MOI of 0.01 or 0.04. At various time points postinfection, the cells were incubated with 99m Tc-pertechnetate, and images of the plates were acquired. (b) Plate layout for hNIS functional assays. (c) Representative images of the plates acquired at 24 h postinfection at an MOI of 0.01 (top plates) or 0.04 (bottom plates) after incubation with 99m Tc-pertechnetate.
    Figure Legend Snippet: Radio-uptake of 99m Tc-pertechnetate by planar scintigraphy. (a) Experimental overview of in vitro evaluation of the rMERS-CoV/ hNIS virus. Vero E6 cells were infected with rMERS-CoV or rMERS-CoV/ hNIS at an MOI of 0.01 or 0.04. At various time points postinfection, the cells were incubated with 99m Tc-pertechnetate, and images of the plates were acquired. (b) Plate layout for hNIS functional assays. (c) Representative images of the plates acquired at 24 h postinfection at an MOI of 0.01 (top plates) or 0.04 (bottom plates) after incubation with 99m Tc-pertechnetate.

    Techniques Used: In Vitro, Infection, Incubation, Functional Assay

    Retention of hNIS transgene following viral kinetics analysis and serial passage. (a and b) Vero E6 cells were infected with rMERS-CoV/hNIS (a) or parental rMERS-CoV (b) at an MOI of 0.01 or 3 and then collected at 96 h postinfection for RT-PCR. (c) Retention of the hNIS gene following serial passage. RNA was extracted from cells 72 h postinfection followed by RT-PCR at passage 6. A positive-control virus (C+) and uninfected negative-control cells (C−) were used as controls.
    Figure Legend Snippet: Retention of hNIS transgene following viral kinetics analysis and serial passage. (a and b) Vero E6 cells were infected with rMERS-CoV/hNIS (a) or parental rMERS-CoV (b) at an MOI of 0.01 or 3 and then collected at 96 h postinfection for RT-PCR. (c) Retention of the hNIS gene following serial passage. RNA was extracted from cells 72 h postinfection followed by RT-PCR at passage 6. A positive-control virus (C+) and uninfected negative-control cells (C−) were used as controls.

    Techniques Used: Infection, Reverse Transcription Polymerase Chain Reaction, Positive Control, Negative Control

    Kinetics of rMERS-CoV/ hNIS and parental rMERS-CoV replication in Vero E6 cells. (a and b) Multistep (a) and one-step (b) growth curves of Vero E6 cells infected with rMERS-CoV (Parental) and rMERS-CoV/ hNIS ( hNIS ). Quantification of the release of infectious virus at the indicated time points (hours postexposure) was determined by plaque assays. Each data point represents the mean ± standard deviation (SD) (error bar) averaged from three independent experiments. (c and d) Cytopathology of rMERS-CoV and rMERS-CoV/ hNIS in Vero E6 cells. The cells were infected with either rMERS-CoV or rMERS-CoV/ hNIS at an MOI of 0.01 (c) or 3 (d) and analyzed by light microscopy at the indicated time points. Photomicrographs were taken using a 40× objective.
    Figure Legend Snippet: Kinetics of rMERS-CoV/ hNIS and parental rMERS-CoV replication in Vero E6 cells. (a and b) Multistep (a) and one-step (b) growth curves of Vero E6 cells infected with rMERS-CoV (Parental) and rMERS-CoV/ hNIS ( hNIS ). Quantification of the release of infectious virus at the indicated time points (hours postexposure) was determined by plaque assays. Each data point represents the mean ± standard deviation (SD) (error bar) averaged from three independent experiments. (c and d) Cytopathology of rMERS-CoV and rMERS-CoV/ hNIS in Vero E6 cells. The cells were infected with either rMERS-CoV or rMERS-CoV/ hNIS at an MOI of 0.01 (c) or 3 (d) and analyzed by light microscopy at the indicated time points. Photomicrographs were taken using a 40× objective.

    Techniques Used: Infection, Standard Deviation, Light Microscopy

    36) Product Images from "Ribavirin Causes Error Catastrophe during Hantaan Virus Replication"

    Article Title: Ribavirin Causes Error Catastrophe during Hantaan Virus Replication

    Journal: Journal of Virology

    doi: 10.1128/JVI.77.1.481-488.2003

    Effect of ribavirin on the yield of infectious HTNV. Supernatants were harvested from HTNV-infected cells at MOIs of 0.01 (A) and 0.1 (B) for the various treatments, i.e., 0 (•) or 24 (▪) μg of ribavirin, and then assayed for plaque formation on monolayers of Vero E6 cells.
    Figure Legend Snippet: Effect of ribavirin on the yield of infectious HTNV. Supernatants were harvested from HTNV-infected cells at MOIs of 0.01 (A) and 0.1 (B) for the various treatments, i.e., 0 (•) or 24 (▪) μg of ribavirin, and then assayed for plaque formation on monolayers of Vero E6 cells.

    Techniques Used: Infection

    37) Product Images from "In Vitro and in Vivo Evaluation of Mutations in the NS Region of Lineage 2 West Nile Virus Associated with Neuroinvasiveness in a Mammalian Model"

    Article Title: In Vitro and in Vivo Evaluation of Mutations in the NS Region of Lineage 2 West Nile Virus Associated with Neuroinvasiveness in a Mammalian Model

    Journal: Viruses

    doi: 10.3390/v8020049

    Growth kinetics of infectious virus of the wild type (WT) virus and mutant viruses after triplicate infection of Vero E6 cells at an MOI of 0.1. The titres are given as the mean (log10 TCID 50 /mL); error bars represent standard deviation.
    Figure Legend Snippet: Growth kinetics of infectious virus of the wild type (WT) virus and mutant viruses after triplicate infection of Vero E6 cells at an MOI of 0.1. The titres are given as the mean (log10 TCID 50 /mL); error bars represent standard deviation.

    Techniques Used: Mutagenesis, Infection, Standard Deviation

    Quantification of ( a ) extracellular positive strand RNA ( b ) intracellular positive strand RNA, and ( c ) negative strand RNA for WT and NS1 mutant after triplicate infection of Vero E6 cells at an MOI of 0.1. Copy numbers are given as the mean of two independent experiments (log10 TCID 50 /mL); error bars represent standard deviation.
    Figure Legend Snippet: Quantification of ( a ) extracellular positive strand RNA ( b ) intracellular positive strand RNA, and ( c ) negative strand RNA for WT and NS1 mutant after triplicate infection of Vero E6 cells at an MOI of 0.1. Copy numbers are given as the mean of two independent experiments (log10 TCID 50 /mL); error bars represent standard deviation.

    Techniques Used: Mutagenesis, Infection, Standard Deviation

    38) Product Images from "Monocytes and macrophages, targets of SARS-CoV-2: the clue for Covid-19 immunoparalysis"

    Article Title: Monocytes and macrophages, targets of SARS-CoV-2: the clue for Covid-19 immunoparalysis

    Journal: bioRxiv

    doi: 10.1101/2020.09.17.300996

    SARS-CoV-2 infects monocytes and macrophages and stimulates cytokine release. Vero E6 cells, monocytes and monocyte-derived macrophages were infected with SARS-CoV-2 IHU-MI3 strain (0.1 MOI) for 24 or 48 hours. ( A ) SARS-CoV-2 quantification was evaluated by RT-PCR, expressed as Ct values and observed in red in infected cells, with the nucleus in blue and F-actin in green (n = 3). Pictures were acquired using a confocal microscope (63x). **** P
    Figure Legend Snippet: SARS-CoV-2 infects monocytes and macrophages and stimulates cytokine release. Vero E6 cells, monocytes and monocyte-derived macrophages were infected with SARS-CoV-2 IHU-MI3 strain (0.1 MOI) for 24 or 48 hours. ( A ) SARS-CoV-2 quantification was evaluated by RT-PCR, expressed as Ct values and observed in red in infected cells, with the nucleus in blue and F-actin in green (n = 3). Pictures were acquired using a confocal microscope (63x). **** P

    Techniques Used: Derivative Assay, Infection, Reverse Transcription Polymerase Chain Reaction, Microscopy

    39) Product Images from "Natural Products Isolated from Oriental Medicinal Herbs Inactivate Zika Virus"

    Article Title: Natural Products Isolated from Oriental Medicinal Herbs Inactivate Zika Virus

    Journal: Viruses

    doi: 10.3390/v11010049

    Pre-treatment of cells with emodin and berberine. Graphic of relative foci number after treatment of Vero E6 cells with berberine or emodin. Cells treated with drug diluent were used as negative control. Results are the means (±SD) from three independent experiments and are expressed as relative values compared to untreated cells. *** p
    Figure Legend Snippet: Pre-treatment of cells with emodin and berberine. Graphic of relative foci number after treatment of Vero E6 cells with berberine or emodin. Cells treated with drug diluent were used as negative control. Results are the means (±SD) from three independent experiments and are expressed as relative values compared to untreated cells. *** p

    Techniques Used: Negative Control

    Viability of emodin- and berberine-treated Vero E6 cells. Cells were incubated with different concentrations of emodin ( A ) or berberine ( B ) and cell viability was evaluated after 24 h, 48 h, and 72 h.
    Figure Legend Snippet: Viability of emodin- and berberine-treated Vero E6 cells. Cells were incubated with different concentrations of emodin ( A ) or berberine ( B ) and cell viability was evaluated after 24 h, 48 h, and 72 h.

    Techniques Used: Incubation

    40) Product Images from "Dynein-Dependent Transport of the Hantaan Virus Nucleocapsid Protein to the Endoplasmic Reticulum-Golgi Intermediate Compartment ▿"

    Article Title: Dynein-Dependent Transport of the Hantaan Virus Nucleocapsid Protein to the Endoplasmic Reticulum-Golgi Intermediate Compartment ▿

    Journal: Journal of Virology

    doi: 10.1128/JVI.00418-07

    Distribution of N in Vero E6 cells and redistribution upon treatment with CytD, BFA, or NOC. Vero E6 cells were transfected with pcHTNVN, which expresses N, and after 18 h the cells were treated with a mock vector (A), CytD (B), BFA (C), or NOC (D). Slides
    Figure Legend Snippet: Distribution of N in Vero E6 cells and redistribution upon treatment with CytD, BFA, or NOC. Vero E6 cells were transfected with pcHTNVN, which expresses N, and after 18 h the cells were treated with a mock vector (A), CytD (B), BFA (C), or NOC (D). Slides

    Techniques Used: Transfection, Plasmid Preparation

    Redistribution of N in HTNV-infected Vero E6 cells and redistribution upon treatment with CytD, BFA, or NOC. Vero E6 cells were infected with HTNV at an MOI of 0.1, and after 3 days the cells were treated for 1 h with a mock vector (A), CytD (B), BFA
    Figure Legend Snippet: Redistribution of N in HTNV-infected Vero E6 cells and redistribution upon treatment with CytD, BFA, or NOC. Vero E6 cells were infected with HTNV at an MOI of 0.1, and after 3 days the cells were treated for 1 h with a mock vector (A), CytD (B), BFA

    Techniques Used: Infection, Plasmid Preparation

    Colocalization of HTNV N with ERGIC-53 and redistribution of N with BFA. Vero E6 cells were infected with HTNV at an MOI of 0.1, and after 3 days slides were acetone fixed (except for Mann II staining, in which case paraformaldehyde was used). Prior to
    Figure Legend Snippet: Colocalization of HTNV N with ERGIC-53 and redistribution of N with BFA. Vero E6 cells were infected with HTNV at an MOI of 0.1, and after 3 days slides were acetone fixed (except for Mann II staining, in which case paraformaldehyde was used). Prior to

    Techniques Used: Infection, Staining

    Relationship of N and vimentin in HTNV-infected and pcHTNVN-transfected Vero E6 cells. Vero E6 cells were infected with HTNV at an MOI of 0.1 (A) or were transfected with pcHTNVN (B), and after 5 days or 24 h, respectively, slides were acetone fixed and
    Figure Legend Snippet: Relationship of N and vimentin in HTNV-infected and pcHTNVN-transfected Vero E6 cells. Vero E6 cells were infected with HTNV at an MOI of 0.1 (A) or were transfected with pcHTNVN (B), and after 5 days or 24 h, respectively, slides were acetone fixed and

    Techniques Used: Infection, Transfection

    Temporal distribution of N in Vero E6 cells infected with HTNV or transfected with plasmid expressing N. (A) To ensure maximal infection, Vero E6 cells were infected with HTNV at an MOI of 5.0 and were examined for the presence of N at 4, 12, 24, and
    Figure Legend Snippet: Temporal distribution of N in Vero E6 cells infected with HTNV or transfected with plasmid expressing N. (A) To ensure maximal infection, Vero E6 cells were infected with HTNV at an MOI of 5.0 and were examined for the presence of N at 4, 12, 24, and

    Techniques Used: Infection, Transfection, Plasmid Preparation, Expressing

    Colocalization studies of HTNV N in Vero E6 cells expressing N alone, as well as redistribution of N with BFA, with various subcellular markers. Vero E6 cells were transfected with pcHTNVN, which expresses N alone, and after 18 h, the cells were fixed
    Figure Legend Snippet: Colocalization studies of HTNV N in Vero E6 cells expressing N alone, as well as redistribution of N with BFA, with various subcellular markers. Vero E6 cells were transfected with pcHTNVN, which expresses N alone, and after 18 h, the cells were fixed

    Techniques Used: Expressing, Transfection

    Association of HTNV N with membrane fractions. Vero E6 cells were transfected with pcHTNVN, and after 18 h they were subjected to membrane floatation (A) or subcellular fractionation (B). Fractions were subjected to Western blotting and were probed with
    Figure Legend Snippet: Association of HTNV N with membrane fractions. Vero E6 cells were transfected with pcHTNVN, and after 18 h they were subjected to membrane floatation (A) or subcellular fractionation (B). Fractions were subjected to Western blotting and were probed with

    Techniques Used: Transfection, Fractionation, Western Blot

    Overexpression of dynamitin abrogated accumulation of N in the perinuclear region. Vero E6 cells were cotransfected with p50-GFP (green) and pcHTNVN (red). (A) Vero E6 cells infected with HTNV are shown, either mock treated (left panel), treated for 1
    Figure Legend Snippet: Overexpression of dynamitin abrogated accumulation of N in the perinuclear region. Vero E6 cells were cotransfected with p50-GFP (green) and pcHTNVN (red). (A) Vero E6 cells infected with HTNV are shown, either mock treated (left panel), treated for 1

    Techniques Used: Over Expression, Infection

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    Article Snippet: .. Human ( Homo sapiens ) adenocarcinoma alveolar basal epithelial A549 (American Type Culture Collection [ATCC], Manassas, VA; #CCL-185) cells, epithelial Hela cells (ATCC, #CCL-2), and hepatocarcinoma Huh-7 cells (a kind gift from Hideki Ebihara, Rocky Mountain Laboratories, Hamilton, MT, USA), and grivet ( Chlorocebus aethiops ) kidney epithelial Vero cells (ATCC, #CCL-81), and Vero E6 cells (ATCC, #CRL-1586) were grown in Gibco Dulbecco’s modified Eagle’s medium (DMEM) (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS, Sigma-Aldrich, St. Louis, MO, USA). .. Syrian golden hamster ( Mesocricetus auratus ) kidney BHK-21 fibroblasts (ATCC, #CCL-10) were grown in Gibco DMEM supplemented with 10% FBS and 5% tryptose phosphate broth (TPB, Thermo Fisher Scientific).

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    ATCC vero e6 cells
    siRNA No. 14 inhibition of SARS-CoV-2 cytopathicity in <t>Vero</t> E6 cells (CPE assay and EC 50 ). Vero E6 cells were infected with SARS-CoV-2 and incubated for 2 days. (a) Mock-siRNA (100 nM). (b) 5 nM. (c) 10 nM. (d) 20 nM. (e) 30 nM. (f) 40 nM. (g) 50 nM. (h) 60 nM. (i) 70 nM. (j) 80 nM. (k) 90 nM. (l) 100 nM. (m) EC 50 of siRNA No. 14 using qRT-PCR.
    Vero E6 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    siRNA No. 14 inhibition of SARS-CoV-2 cytopathicity in Vero E6 cells (CPE assay and EC 50 ). Vero E6 cells were infected with SARS-CoV-2 and incubated for 2 days. (a) Mock-siRNA (100 nM). (b) 5 nM. (c) 10 nM. (d) 20 nM. (e) 30 nM. (f) 40 nM. (g) 50 nM. (h) 60 nM. (i) 70 nM. (j) 80 nM. (k) 90 nM. (l) 100 nM. (m) EC 50 of siRNA No. 14 using qRT-PCR.

    Journal: bioRxiv

    Article Title: A Small interfering RNA lead targeting RNA-dependent RNA-polymerase effectively inhibit the SARS-CoV-2 infection in Golden Syrian hamster and Rhesus macaque

    doi: 10.1101/2020.07.07.190967

    Figure Lengend Snippet: siRNA No. 14 inhibition of SARS-CoV-2 cytopathicity in Vero E6 cells (CPE assay and EC 50 ). Vero E6 cells were infected with SARS-CoV-2 and incubated for 2 days. (a) Mock-siRNA (100 nM). (b) 5 nM. (c) 10 nM. (d) 20 nM. (e) 30 nM. (f) 40 nM. (g) 50 nM. (h) 60 nM. (i) 70 nM. (j) 80 nM. (k) 90 nM. (l) 100 nM. (m) EC 50 of siRNA No. 14 using qRT-PCR.

    Article Snippet: In vitro efficacy test using Vero E6 cells (plaque assay) To assess viral titers, a plaque assay was performed using Vero E6 cells in 6-well culture plates.

    Techniques: Inhibition, Infection, Incubation, Quantitative RT-PCR

    Radio-uptake of 99m Tc-pertechnetate by planar scintigraphy. (a) Experimental overview of in vitro evaluation of the rMERS-CoV/ hNIS virus. Vero E6 cells were infected with rMERS-CoV or rMERS-CoV/ hNIS at an MOI of 0.01 or 0.04. At various time points postinfection, the cells were incubated with 99m Tc-pertechnetate, and images of the plates were acquired. (b) Plate layout for hNIS functional assays. (c) Representative images of the plates acquired at 24 h postinfection at an MOI of 0.01 (top plates) or 0.04 (bottom plates) after incubation with 99m Tc-pertechnetate.

    Journal: mSphere

    Article Title: The Human Sodium Iodide Symporter as a Reporter Gene for Studying Middle East Respiratory Syndrome Coronavirus Pathogenesis

    doi: 10.1128/mSphere.00540-18

    Figure Lengend Snippet: Radio-uptake of 99m Tc-pertechnetate by planar scintigraphy. (a) Experimental overview of in vitro evaluation of the rMERS-CoV/ hNIS virus. Vero E6 cells were infected with rMERS-CoV or rMERS-CoV/ hNIS at an MOI of 0.01 or 0.04. At various time points postinfection, the cells were incubated with 99m Tc-pertechnetate, and images of the plates were acquired. (b) Plate layout for hNIS functional assays. (c) Representative images of the plates acquired at 24 h postinfection at an MOI of 0.01 (top plates) or 0.04 (bottom plates) after incubation with 99m Tc-pertechnetate.

    Article Snippet: Vero E6 cells (ATCC CRL-1586) were maintained in Dulbecco’s modified Eagle medium (DMEM) (Lonza) supplemented with 5% fetal bovine serum (FBS) and incubated at 37°C and 5% CO2 without antibiotics or antimycotics.

    Techniques: In Vitro, Infection, Incubation, Functional Assay

    Retention of hNIS transgene following viral kinetics analysis and serial passage. (a and b) Vero E6 cells were infected with rMERS-CoV/hNIS (a) or parental rMERS-CoV (b) at an MOI of 0.01 or 3 and then collected at 96 h postinfection for RT-PCR. (c) Retention of the hNIS gene following serial passage. RNA was extracted from cells 72 h postinfection followed by RT-PCR at passage 6. A positive-control virus (C+) and uninfected negative-control cells (C−) were used as controls.

    Journal: mSphere

    Article Title: The Human Sodium Iodide Symporter as a Reporter Gene for Studying Middle East Respiratory Syndrome Coronavirus Pathogenesis

    doi: 10.1128/mSphere.00540-18

    Figure Lengend Snippet: Retention of hNIS transgene following viral kinetics analysis and serial passage. (a and b) Vero E6 cells were infected with rMERS-CoV/hNIS (a) or parental rMERS-CoV (b) at an MOI of 0.01 or 3 and then collected at 96 h postinfection for RT-PCR. (c) Retention of the hNIS gene following serial passage. RNA was extracted from cells 72 h postinfection followed by RT-PCR at passage 6. A positive-control virus (C+) and uninfected negative-control cells (C−) were used as controls.

    Article Snippet: Vero E6 cells (ATCC CRL-1586) were maintained in Dulbecco’s modified Eagle medium (DMEM) (Lonza) supplemented with 5% fetal bovine serum (FBS) and incubated at 37°C and 5% CO2 without antibiotics or antimycotics.

    Techniques: Infection, Reverse Transcription Polymerase Chain Reaction, Positive Control, Negative Control

    Kinetics of rMERS-CoV/ hNIS and parental rMERS-CoV replication in Vero E6 cells. (a and b) Multistep (a) and one-step (b) growth curves of Vero E6 cells infected with rMERS-CoV (Parental) and rMERS-CoV/ hNIS ( hNIS ). Quantification of the release of infectious virus at the indicated time points (hours postexposure) was determined by plaque assays. Each data point represents the mean ± standard deviation (SD) (error bar) averaged from three independent experiments. (c and d) Cytopathology of rMERS-CoV and rMERS-CoV/ hNIS in Vero E6 cells. The cells were infected with either rMERS-CoV or rMERS-CoV/ hNIS at an MOI of 0.01 (c) or 3 (d) and analyzed by light microscopy at the indicated time points. Photomicrographs were taken using a 40× objective.

    Journal: mSphere

    Article Title: The Human Sodium Iodide Symporter as a Reporter Gene for Studying Middle East Respiratory Syndrome Coronavirus Pathogenesis

    doi: 10.1128/mSphere.00540-18

    Figure Lengend Snippet: Kinetics of rMERS-CoV/ hNIS and parental rMERS-CoV replication in Vero E6 cells. (a and b) Multistep (a) and one-step (b) growth curves of Vero E6 cells infected with rMERS-CoV (Parental) and rMERS-CoV/ hNIS ( hNIS ). Quantification of the release of infectious virus at the indicated time points (hours postexposure) was determined by plaque assays. Each data point represents the mean ± standard deviation (SD) (error bar) averaged from three independent experiments. (c and d) Cytopathology of rMERS-CoV and rMERS-CoV/ hNIS in Vero E6 cells. The cells were infected with either rMERS-CoV or rMERS-CoV/ hNIS at an MOI of 0.01 (c) or 3 (d) and analyzed by light microscopy at the indicated time points. Photomicrographs were taken using a 40× objective.

    Article Snippet: Vero E6 cells (ATCC CRL-1586) were maintained in Dulbecco’s modified Eagle medium (DMEM) (Lonza) supplemented with 5% fetal bovine serum (FBS) and incubated at 37°C and 5% CO2 without antibiotics or antimycotics.

    Techniques: Infection, Standard Deviation, Light Microscopy